Polymerase Chain Reaction
In vitro method for producing large amounts of specific DNA or RNA fragments of defined length and sequence from small amounts of short oligonucleotide flanking sequences (primers). The essential steps include thermal denaturation of the double-stranded target molecules, annealing of the primers to their complementary sequences, and extension of the annealed primers by enzymatic synthesis with DNA polymerase. The reaction is efficient, specific, and extremely sensitive. Uses for the reaction include disease diagnosis, detection of difficult-to-isolate pathogens, mutation analysis, genetic testing, DNA sequencing, and analyzing evolutionary relationships.
Reverse Transcriptase Polymerase Chain Reaction
Molecular Sequence Data
Descriptions of specific amino acid, carbohydrate, or nucleotide sequences which have appeared in the published literature and/or are deposited in and maintained by databanks such as GENBANK, European Molecular Biology Laboratory (EMBL), National Biomedical Research Foundation (NBRF), or other sequence repositories.
DNA-Directed DNA Polymerase
DNA-dependent DNA polymerases found in bacteria, animal and plant cells. During the replication process, these enzymes catalyze the addition of deoxyribonucleotide residues to the end of a DNA strand in the presence of DNA as template-primer. They also possess exonuclease activity and therefore function in DNA repair.
RNA sequences that serve as templates for protein synthesis. Bacterial mRNAs are generally primary transcripts in that they do not require post-transcriptional processing. Eukaryotic mRNA is synthesized in the nucleus and must be exported to the cytoplasm for translation. Most eukaryotic mRNAs have a sequence of polyadenylic acid at the 3' end, referred to as the poly(A) tail. The function of this tail is not known for certain, but it may play a role in the export of mature mRNA from the nucleus as well as in helping stabilize some mRNA molecules by retarding their degradation in the cytoplasm.
Real-Time Polymerase Chain Reaction
RNA Polymerase II
Sensitivity and Specificity
Amino Acid Sequence
A deoxyribonucleotide polymer that is the primary genetic material of all cells. Eukaryotic and prokaryotic organisms normally contain DNA in a double-stranded state, yet several important biological processes transiently involve single-stranded regions. DNA, which consists of a polysugar-phosphate backbone possessing projections of purines (adenine and guanine) and pyrimidines (thymine and cytosine), forms a double helix that is held together by hydrogen bonds between these purines and pyrimidines (adenine to thymine and guanine to cytosine).
DNA-Directed RNA Polymerases
Enzymes that catalyze DNA template-directed extension of the 3'-end of an RNA strand one nucleotide at a time. They can initiate a chain de novo. In eukaryotes, three forms of the enzyme have been distinguished on the basis of sensitivity to alpha-amanitin, and the type of RNA synthesized. (From Enzyme Nomenclature, 1992).
DNA Polymerase I
A DNA-dependent DNA polymerase characterized in prokaryotes and may be present in higher organisms. It has both 3'-5' and 5'-3' exonuclease activity, but cannot use native double-stranded DNA as template-primer. It is not inhibited by sulfhydryl reagents and is active in both DNA synthesis and repair. EC 220.127.116.11.
Synthetic or natural oligonucleotides used in hybridization studies in order to identify and study specific nucleic acid fragments, e.g., DNA segments near or within a specific gene locus or gene. The probe hybridizes with a specific mRNA, if present. Conventional techniques used for testing for the hybridization product include dot blot assays, Southern blot assays, and DNA:RNA hybrid-specific antibody tests. Conventional labels for the probe include the radioisotope labels 32P and 125I and the chemical label biotin.
DNA Polymerase II
A DNA-dependent DNA polymerase characterized in E. coli and other lower organisms. It may be present in higher organisms and has an intrinsic molecular activity only 5% of that of DNA Polymerase I. This polymerase has 3'-5' exonuclease activity, is effective only on duplex DNA with gaps or single-strand ends of less than 100 nucleotides as template, and is inhibited by sulfhydryl reagents. EC 18.104.22.168.
Multiplex Polymerase Chain Reaction
Polymorphism, Restriction Fragment Length
DNA Polymerase III
A DNA-dependent DNA polymerase characterized in E. coli and other lower organisms but may be present in higher organisms. Use also for a more complex form of DNA polymerase III designated as DNA polymerase III* or pol III* which is 15 times more active biologically than DNA polymerase I in the synthesis of DNA. This polymerase has both 3'-5' and 5'-3' exonuclease activities, is inhibited by sulfhydryl reagents, and has the same template-primer dependence as pol II. EC 22.214.171.124.
Sequence Analysis, DNA
Electrophoresis, Agar Gel
A selective increase in the number of copies of a gene coding for a specific protein without a proportional increase in other genes. It occurs naturally via the excision of a copy of the repeating sequence from the chromosome and its extrachromosomal replication in a plasmid, or via the production of an RNA transcript of the entire repeating sequence of ribosomal RNA followed by the reverse transcription of the molecule to produce an additional copy of the original DNA sequence. Laboratory techniques have been introduced for inducing disproportional replication by unequal crossing over, uptake of DNA from lysed cells, or generation of extrachromosomal sequences from rolling circle replication.
The regular and simultaneous occurrence in a single interbreeding population of two or more discontinuous genotypes. The concept includes differences in genotypes ranging in size from a single nucleotide site (POLYMORPHISM, SINGLE NUCLEOTIDE) to large nucleotide sequences visible at a chromosomal level.
Species- or subspecies-specific DNA (including COMPLEMENTARY DNA; conserved genes, whole chromosomes, or whole genomes) used in hybridization studies in order to identify microorganisms, to measure DNA-DNA homologies, to group subspecies, etc. The DNA probe hybridizes with a specific mRNA, if present. Conventional techniques used for testing for the hybridization product include dot blot assays, Southern blot assays, and DNA:RNA hybrid-specific antibody tests. Conventional labels for the DNA probe include the radioisotope labels 32P and 125I and the chemical label biotin. The use of DNA probes provides a specific, sensitive, rapid, and inexpensive replacement for cell culture techniques for diagnosing infections.
RNA Polymerase I
RNA Polymerase III
A DNA-dependent RNA polymerase present in bacterial, plant, and animal cells. It functions in the nucleoplasmic structure where it transcribes DNA into RNA. It has specific requirements for cations and salt and has shown an intermediate sensitivity to alpha-amanitin in comparison to RNA polymerase I and II. EC 126.96.36.199.
Immunoglobulin Heavy Chains
A species of gram-negative, facultatively anaerobic, rod-shaped bacteria (GRAM-NEGATIVE FACULTATIVELY ANAEROBIC RODS) commonly found in the lower part of the intestine of warm-blooded animals. It is usually nonpathogenic, but some strains are known to produce DIARRHEA and pyogenic infections. Pathogenic strains (virotypes) are classified by their specific pathogenic mechanisms such as toxins (ENTEROTOXIGENIC ESCHERICHIA COLI), etc.
RNA-Directed DNA Polymerase
DNA Polymerase beta
Promoter Regions, Genetic
Sequence Homology, Nucleic Acid
Sequence Homology, Amino Acid
Nucleic Acid Hybridization
Widely used technique which exploits the ability of complementary sequences in single-stranded DNAs or RNAs to pair with each other to form a double helix. Hybridization can take place between two complimentary DNA sequences, between a single-stranded DNA and a complementary RNA, or between two RNA sequences. The technique is used to detect and isolate specific sequences, measure homology, or define other characteristics of one or both strands. (Kendrew, Encyclopedia of Molecular Biology, 1994, p503)
In Situ Hybridization
Gene Expression Regulation
A polynucleotide consisting essentially of chains with a repeating backbone of phosphate and ribose units to which nitrogenous bases are attached. RNA is unique among biological macromolecules in that it can encode genetic information, serve as an abundant structural component of cells, and also possesses catalytic activity. (Rieger et al., Glossary of Genetics: Classical and Molecular, 5th ed)
The arrangement of two or more amino acid or base sequences from an organism or organisms in such a way as to align areas of the sequences sharing common properties. The degree of relatedness or homology between the sequences is predicted computationally or statistically based on weights assigned to the elements aligned between the sequences. This in turn can serve as a potential indicator of the genetic relatedness between the organisms.
The restriction of a characteristic behavior, anatomical structure or physical system, such as immune response; metabolic response, or gene or gene variant to the members of one species. It refers to that property which differentiates one species from another but it is also used for phylogenetic levels higher or lower than the species.
Gene Expression Profiling
Studies which start with the identification of persons with a disease of interest and a control (comparison, referent) group without the disease. The relationship of an attribute to the disease is examined by comparing diseased and non-diseased persons with regard to the frequency or levels of the attribute in each group.
Evaluation Studies as Topic
Immunoglobulin Light Chains
Polymorphism, Single-Stranded Conformational
Variation in a population's DNA sequence that is detected by determining alterations in the conformation of denatured DNA fragments. Denatured DNA fragments are allowed to renature under conditions that prevent the formation of double-stranded DNA and allow secondary structure to form in single stranded fragments. These fragments are then run through polyacrylamide gels to detect variations in the secondary structure that is manifested as an alteration in migration through the gels.
Nucleic Acid Amplification Techniques
Enzyme-Linked Immunosorbent Assay
An immunoassay utilizing an antibody labeled with an enzyme marker such as horseradish peroxidase. While either the enzyme or the antibody is bound to an immunosorbent substrate, they both retain their biologic activity; the change in enzyme activity as a result of the enzyme-antibody-antigen reaction is proportional to the concentration of the antigen and can be measured spectrophotometrically or with the naked eye. Many variations of the method have been developed.
Tumor Virus Infections
A family of small, non-enveloped DNA viruses infecting birds and most mammals, especially humans. They are grouped into multiple genera, but the viruses are highly host-species specific and tissue-restricted. They are commonly divided into hundreds of papillomavirus "types", each with specific gene function and gene control regions, despite sequence homology. Human papillomaviruses are found in the genera ALPHAPAPILLOMAVIRUS; BETAPAPILLOMAVIRUS; GAMMAPAPILLOMAVIRUS; and MUPAPILLOMAVIRUS.
Reproducibility of Results
The statistical reproducibility of measurements (often in a clinical context), including the testing of instrumentation or techniques to obtain reproducible results. The concept includes reproducibility of physiological measurements, which may be used to develop rules to assess probability or prognosis, or response to a stimulus; reproducibility of occurrence of a condition; and reproducibility of experimental results.
Oligonucleotide Array Sequence Analysis
Electrophoresis, Polyacrylamide Gel
Tumor Cells, Cultured
Genetic Predisposition to Disease
Proteins whose abnormal expression (gain or loss) are associated with the development, growth, or progression of NEOPLASMS. Some neoplasm proteins are tumor antigens (ANTIGENS, NEOPLASM), i.e. they induce an immune reaction to their tumor. Many neoplasm proteins have been characterized and are used as tumor markers (BIOMARKERS, TUMOR) when they are detectable in cells and body fluids as monitors for the presence or growth of tumors. Abnormal expression of ONCOGENE PROTEINS is involved in neoplastic transformation, whereas the loss of expression of TUMOR SUPPRESSOR PROTEINS is involved with the loss of growth control and progression of the neoplasm.
Gene Expression Regulation, Neoplastic
Gene Rearrangement, B-Lymphocyte, Heavy Chain
Any of various animals that constitute the family Suidae and comprise stout-bodied, short-legged omnivorous mammals with thick skin, usually covered with coarse bristles, a rather long mobile snout, and small tail. Included are the genera Babyrousa, Phacochoerus (wart hogs), and Sus, the latter containing the domestic pig (see SUS SCROFA).
Ligase Chain Reaction
A DNA amplification technique based upon the ligation of OLIGONUCLEOTIDE PROBES. The probes are designed to exactly match two adjacent sequences of a specific target DNA. The chain reaction is repeated in three steps in the presence of excess probe: (1) heat denaturation of double-stranded DNA, (2) annealing of probes to target DNA, and (3) joining of the probes by thermostable DNA ligase. After the reaction is repeated for 20-30 cycles the production of ligated probe is measured.
RNA, Ribosomal, 16S
Chromosomes, Human, Pair 14
A genus of the family HERPESVIRIDAE, subfamily BETAHERPESVIRINAE, infecting the salivary glands, liver, spleen, lungs, eyes, and other organs, in which they produce characteristically enlarged cells with intranuclear inclusions. Infection with Cytomegalovirus is also seen as an opportunistic infection in AIDS.
Technique using an instrument system for making, processing, and displaying one or more measurements on individual cells obtained from a cell suspension. Cells are usually stained with one or more fluorescent dyes specific to cell components of interest, e.g., DNA, and fluorescence of each cell is measured as it rapidly transverses the excitation beam (laser or mercury arc lamp). Fluorescence provides a quantitative measure of various biochemical and biophysical properties of the cell, as well as a basis for cell sorting. Other measurable optical parameters include light absorption and light scattering, the latter being applicable to the measurement of cell size, shape, density, granularity, and stain uptake.
Genes encoding the different subunits of the IMMUNOGLOBULINS, for example the IMMUNOGLOBULIN LIGHT CHAIN GENES and the IMMUNOGLOBULIN HEAVY CHAIN GENES. The heavy and light immunoglobulin genes are present as gene segments in the germline cells. The completed genes are created when the segments are shuffled and assembled (B-LYMPHOCYTE GENE REARRANGEMENT) during B-LYMPHOCYTE maturation. The gene segments of the human light and heavy chain germline genes are symbolized V (variable), J (joining) and C (constant). The heavy chain germline genes have an additional segment D (diversity).
Myosin Heavy Chains
Protozoan Infections, Animal
Asian Continental Ancestry Group
A set of three nucleotides in a protein coding sequence that specifies individual amino acids or a termination signal (CODON, TERMINATOR). Most codons are universal, but some organisms do not produce the transfer RNAs (RNA, TRANSFER) complementary to all codons. These codons are referred to as unassigned codons (CODONS, NONSENSE).
Chromosomes, Human, Pair 18
Tumor Markers, Biological
Molecular products metabolized and secreted by neoplastic tissue and characterized biochemically in cells or body fluids. They are indicators of tumor stage and grade as well as useful for monitoring responses to treatment and predicting recurrence. Many chemical groups are represented including hormones, antigens, amino and nucleic acids, enzymes, polyamines, and specific cell membrane proteins and lipids.
Gene Rearrangement, gamma-Chain T-Cell Antigen Receptor
A technique for identifying individuals of a species that is based on the uniqueness of their DNA sequence. Uniqueness is determined by identifying which combination of allelic variations occur in the individual at a statistically relevant number of different loci. In forensic studies, RESTRICTION FRAGMENT LENGTH POLYMORPHISM of multiple, highly polymorphic VNTR LOCI or MICROSATELLITE REPEAT loci are analyzed. The number of loci used for the profile depends on the ALLELE FREQUENCY in the population.
Herpesvirus 6, Human
The type species of ROSEOLOVIRUS isolated from patients with AIDS and other LYMPHOPROLIFERATIVE DISORDERS. It infects and replicates in fresh and established lines of hematopoietic cells and cells of neural origin. It also appears to alter NK cell activity. HHV-6; (HBLV) antibodies are elevated in patients with AIDS, Sjogren's syndrome, sarcoidosis, chronic fatigue syndrome, and certain malignancies. HHV-6 is the cause of EXANTHEMA SUBITUM and has been implicated in encephalitis.
Herpesvirus 4, Human
Repetitive Sequences, Nucleic Acid
Sequences of DNA or RNA that occur in multiple copies. There are several types: INTERSPERSED REPETITIVE SEQUENCES are copies of transposable elements (DNA TRANSPOSABLE ELEMENTS or RETROELEMENTS) dispersed throughout the genome. TERMINAL REPEAT SEQUENCES flank both ends of another sequence, for example, the long terminal repeats (LTRs) on RETROVIRUSES. Variations may be direct repeats, those occurring in the same direction, or inverted repeats, those opposite to each other in direction. TANDEM REPEAT SEQUENCES are copies which lie adjacent to each other, direct or inverted (INVERTED REPEAT SEQUENCES).
Fluorescent Antibody Technique, Direct
Polymorphism, Single Nucleotide
Disease Models, Animal
A group of genetically identical cells all descended from a single common ancestral cell by mitosis in eukaryotes or by binary fission in prokaryotes. Clone cells also include populations of recombinant DNA molecules all carrying the same inserted sequence. (From King & Stansfield, Dictionary of Genetics, 4th ed)
The soft tissue filling the cavities of bones. Bone marrow exists in two types, yellow and red. Yellow marrow is found in the large cavities of large bones and consists mostly of fat cells and a few primitive blood cells. Red marrow is a hematopoietic tissue and is the site of production of erythrocytes and granular leukocytes. Bone marrow is made up of a framework of connective tissue containing branching fibers with the frame being filled with marrow cells.
RNA, Ribosomal, 18S
DNA of kinetoplasts which are specialized MITOCHONDRIA of trypanosomes and related parasitic protozoa within the order KINETOPLASTIDA. Kinetoplast DNA consists of a complex network of numerous catenated rings of two classes; the first being a large number of small DNA duplex rings, called minicircles, approximately 2000 base pairs in length, and the second being several dozen much larger rings, called maxicircles, approximately 37 kb in length.
Gene Expression Regulation, Enzymologic
In Situ Hybridization, Fluorescence
Fluorescent Antibody Technique, Indirect
A form of fluorescent antibody technique commonly used to detect serum antibodies and immune complexes in tissues and microorganisms in specimens from patients with infectious diseases. The technique involves formation of an antigen-antibody complex which is labeled with fluorescein-conjugated anti-immunoglobulin antibody. (From Bennington, Saunders Dictionary & Encyclopedia of Laboratory Medicine and Technology, 1984)
Predictive Value of Tests
In screening and diagnostic tests, the probability that a person with a positive test is a true positive (i.e., has the disease), is referred to as the predictive value of a positive test; whereas, the predictive value of a negative test is the probability that the person with a negative test does not have the disease. Predictive value is related to the sensitivity and specificity of the test.
Family of retrovirus-associated DNA sequences (ras) originally isolated from Harvey (H-ras, Ha-ras, rasH) and Kirsten (K-ras, Ki-ras, rasK) murine sarcoma viruses. Ras genes are widely conserved among animal species and sequences corresponding to both H-ras and K-ras genes have been detected in human, avian, murine, and non-vertebrate genomes. The closely related N-ras gene has been detected in human neuroblastoma and sarcoma cell lines. All genes of the family have a similar exon-intron structure and each encodes a p21 protein.
Detailed methylation analysis of the glutathione S-transferase pi (GSTP1) gene in prostate cancer. (1/67158)Glutathione-S-Transferases (GSTs) comprise a family of isoenzymes that provide protection to mammalian cells against electrophilic metabolites of carcinogens and reactive oxygen species. Previous studies have shown that the CpG-rich promoter region of the pi-class gene GSTP1 is methylated at single restriction sites in the majority of prostate cancers. In order to understand the nature of abnormal methylation of the GSTP1 gene in prostate cancer we undertook a detailed analysis of methylation at 131 CpG sites spanning the promoter and body of the gene. Our results show that DNA methylation is not confined to specific CpG sites in the promoter region of the GSTP1 gene but is extensive throughout the CpG island in prostate cancer cells. Furthermore we found that both alleles are abnormally methylated in this region. In normal prostate tissue, the entire CpG island was unmethylated, but extensive methylation was found outside the island in the body of the gene. Loss of GSTP1 expression correlated with DNA methylation of the CpG island in both prostate cancer cell lines and cancer tissues whereas methylation outside the CpG island in normal prostate tissue appeared to have no effect on gene expression. (+info)
Differential stability of the DNA-activated protein kinase catalytic subunit mRNA in human glioma cells. (2/67158)DNA-dependent protein kinase (DNA-PK) functions in double-strand break repair and immunoglobulin [V(D)J] recombination. We previously established a radiation-sensitive human cell line, M059J, derived from a malignant glioma, which lacks the catalytic subunit (DNA-PKcs) of the DNA-PK multiprotein complex. Although previous Northern blot analysis failed to detect the DNA-PKcs transcript in these cells, we show here through quantitative studies that the transcript is present, albeit at greatly reduced (approximately 20x) levels. Sequencing revealed no genetic alteration in either the promoter region, the kinase domain, or the 3' untranslated region of the DNA-PKcs gene to account for the reduced transcript levels. Nuclear run-on transcription assays indicated that the rate of DNA-PKcs transcription in M059J and DNA-PKcs proficient cell lines was similar, but the stability of the DNA-PKcs message in the M059J cell line was drastically (approximately 20x) reduced. Furthermore, M059J cells lack an alternately spliced DNA-PKcs transcript that accounts for a minor (5-20%) proportion of the DNA-PKcs message in all other cell lines tested. Thus, alterations in DNA-PKcs mRNA stability and/or the lack of the alternate mRNA may result in the loss of DNA-PKcs activity. This finding has important implications as DNA-PKcs activity is essential to cells repairing damage induced by radiation or radiomimetric agents. (+info)
Hybrid capture II, a new sensitive test for human papillomavirus detection. Comparison with hybrid capture I and PCR results in cervical lesions. (3/67158)AIM: To test a new assay for the detection of human papillomavirus (HPV) DNA, hybrid capture II (HC II), compared with the previous commercialized hybrid capture I (HC I) and polymerase chain reaction (PCR) results on cervical scrapes from fresh cone excision biopsy samples. METHODS: The three methods were used on cervical scrapes from 42 fresh cone excision biopsy samples. There were nine metaplastic and inflammatory lesions, five low grade lesions, and 28 high grade lesions. PCR was performed using the general primers GP5+/GP6+. The viral load of high risk HPV DNA was estimated by the ratio of relative light units to positive control values in the samples. RESULTS: The sensitivity of HC I for the detection of high grade lesions was 71.4%, while it was 92.8% for HC II and 96.4% for the PCR. Considering only the absence of detectable cervical in situ neoplasia, the specificity was 88.9% for HC I, 66.7% for HC II, and 66.7% for PCR. With HC II, for a ratio of cervical sample to normal control of > 200, the sensitivity for the detection of high grade lesion was only 34.6% with a specificity of 66.7%. CONCLUSIONS: HPV detection with the HC II assay is more sensitive than the previous HC I and represents a more convenient and easier test than PCR for routine use. Nevertheless the viral load estimated with this test cannot be a reliable predictive indicator of high grade lesions. (+info)
Human papillomavirus DNA in adenosquamous carcinoma of the lung. (4/67158)AIM: To investigate the presence of human papillomavirus (HPV) DNA in adenosquamous carcinoma of the lung--which is relatively common in Okinawa but not in mainland Japan--and examine its histological features. METHODS: Of 207 cases where primary lung cancers were surgically removed between January 1995 and June 1997 in Okinawa, 23 were adenosquamous carcinoma. HPV was detected by non-isotopic in situ hybridisation (NISH) and polymerase chain reaction (PCR) amplification with primers specific for E6 and E7 regions of the HPV genome. PCR products were analysed by Southern blotting. Immunohistochemical determination of high molecular weight cytokeratin (HMC) and involucrin was also carried out. RESULTS: 18 cases were positive for HPV DNA by PCR and NISH. HPV types 6, 11, 16, and 18 were found. Seven cases were dual positive for different types of HPV. Using NISH, HPV was also found in the squamous cell components and in neighbouring enlarged adenocarcinoma cells. The HMC and involucrin were demonstrated immunohistochemically in the same areas. CONCLUSIONS: HPV DNA was found in a high proportion (78.3%) of adenosquamous carcinomas in Okinawa, a region where HPV has previously been shown to be prevalent in squamous cell carcinoma of the lung. The adenocarcinoma cells adjacent to the squamous cell carcinoma component were enlarged and positive for HPV, HMC, and involucrin. This is thought to indicate the transition from adenocarcinoma to squamous cell carcinoma. (+info)
Immunohistochemical expression of mdm2 and p21WAF1 in invasive cervical cancer: correlation with p53 protein and high risk HPV infection. (5/67158)AIM: To investigate the immunocytochemical staining pattern of mdm2 and p21WAF1 proteins in invasive cervical cancer and to determine its relation with the expression of p53 and with the high risk HPV infection. METHODS: Immunocytochemistry for p53, mdm2, and p21WAF1 was performed in 31 paraffin embedded sections of invasive cervical cancer. The results were assessed by image analysis, evaluating for each protein the optical density of the immunostained area, scored as percentage of the total nuclear area. The presence of high risk human papillomavirus (HPV) infection was detected by using the polymerase chain reaction. RESULTS: Immunostaining for both mdm2 and p21WAF1 was correlated with p53 expression; however, the correlation between p53 and mdm2 (R = 0.49; p < 0.01) was more significant than between p53 and p21WAF1 (R = 0.31; p < 0.05); the less stringent correlation between p53 and p21WAF1 might reflect the p53 independent mechanisms of p21WAF1 induction. Similar average levels of p53, mdm2, and p21WAF1 immunostaining were found in the presence or absence of high risk HPV-DNA, without significant differences between the two groups. CONCLUSIONS: These data suggest that mdm2 and p21WAF1 proteins are expressed in invasive cervical cancer and that their immunocytochemical staining pattern is not abrogated by the presence of high risk HPV genomic sequences. (+info)
The significance of cagA and vacA subtypes of Helicobacter pylori in the pathogenesis of inflammation and peptic ulceration. (6/67158)AIMS: To assess the significance of cagA and vacA subtypes of Helicobacter pylori in relation to inflammation and density of bacterial colonisation in vivo within a dyspeptic UK population. METHODS: Dyspeptic patients who were Helicobacter pylori positive had antral samples taken for histology and culture. Gastroduodenal pathology was noted. The grade of bacterial density and inflammation was assessed using the Sydney system. Bacterial DNA was extracted and the vacA alleles and the cagA/gene typed using PCR. RESULTS: 120 patients were studied. There was high rate of cagA positive strains in this population. Bacterial density did not correlate with the presence of peptic ulceration. There was a significant association between cagA positive strains and increased inflammation and bacterial density. The vacA s1 type independently correlated with extensive chronic inflammation but there was no association with bacterial density. The vacA m type did not correlate with extent of inflammation or bacterial density. CONCLUSIONS: The results suggest that cagA is important in the pathogenesis of inflammation and peptic ulceration. These findings are in keeping with the hypothesis that cagA acts as a marker for a cag pathogenicity island which encodes several genes involved in inflammation. The vacA s1 allele correlates with inflammation independently of cagA, possibly through its enhanced ability to produce the vacuolating cytotoxin. (+info)
Expression of vascular endothelial growth factor in human oral squamous cell carcinoma: its association with tumour progression and p53 gene status. (7/67158)AIMS: To correlate vascular endothelial growth factor (VEGF) expression in oral squamous cell carcinoma with the clinicopathological characteristics and prognosis; and to assess whether p53 gene status is associated with VEGF expression in human cancers. METHODS: Tumour specimens from 45 patients with oral squamous cell carcinomas were examined. Expression of VEGF was determined using an immunohistochemical method, and a tumour was considered positive when more than 5% of the neoplastic cells showed VEGF immunoreactivity. The p53 gene status was screened using a polymerase chain reaction--single strand conformation polymorphism analysis. RESULTS: VEGF positive staining was detected in 19 (42.2%) of the 45 cases. VEGF immunoreactivity did not correlate with the histological degree of tumour differentiation, clinical stages, or lymph node metastasis. The patients with VEGF positive tumours had a significantly worse prognosis than those with VEGF negative tumours. The five year overall survival rate of the VEGF negative patients was 76.5%, as compared with 48.8% for the VEGF positive patients. No significant association between VEGF expression and the p53 gene status of the tumours was found. CONCLUSIONS: VEGF is a good prognostic indicator of the survival of patients with oral squamous cell carcinoma. The p53 gene status does not seem to be associated with VEGF expression in these cancers. (+info)
The alphaE-catenin gene (CTNNA1) acts as an invasion-suppressor gene in human colon cancer cells. (8/67158)The acquisition of invasiveness is a crucial step in the malignant progression of cancer. In cancers of the colon and of other organs the E-cadherin/catenin complex, which is implicated in homotypic cell-cell adhesion as well as in signal transduction, serves as a powerful inhibitor of invasion. We show here that one allele of the alphaE-catenin (CTNNA1) gene is mutated in the human colon cancer cell family HCT-8, which is identical to HCT-15, DLD-1 and HRT-18. Genetic instability, due to mutations in the HMSH6 (also called GTBP) mismatch repair gene, results in the spontaneous occurrence of invasive variants, all carrying either a mutation or exon skipping in the second alphaE-catenin allele. The alphaE-catenin gene is therefore, an invasion-suppressor gene in accordance with the two-hit model of Knudsen for tumour-suppressor genes. (+info)
A Simple Polymerase Chain Reaction-based Method for the Discrimination of Three Chicken Breeds | Korea Science
A Simple Polymerase Chain Reaction-based Method for the Discrimination of Three Chicken Breeds - Amplified Fragment Length Polymorphism;Breed Discrimination;Chicken;Polymerase Chain Reaction;Nucleotide Polymorphism;
Real‐time polymerase chain reaction tests versus antenatal culture tests for the screening of maternal group B Streptococcus...
Seedat, Farah, Uthman, Olalekan, Robinson, Esther, Cooper, Jennifer, Takwoingi, Yemisi, Kandala, Ngianga-Bakwin, Stranges, Saverio and Taylor-Phillips, Sian (2018) Real‐time polymerase chain reaction tests versus antenatal culture tests for the screening of maternal group B Streptococcus colonisation in labour. Cochrane Database of Systematic Reviews, 2018 (5). CD013016. ISSN 1465-1858 ...
Patente US5538871 - In situ polymerase chain reaction - Google Patentes
Improvements to the in situ polymerase chain reaction (PCR), a process of in vitro enzymatic amplification of specific nucleic acid sequences within the cells where they originate, can be achieved by changing the way that the enzymatic reaction is started. Reaction initiation is delayed until the start of PCR thermal cycling, either by withholding a subset of PCR reagents from the cellular preparation until the preparation has been heated to 50 C. to 80 C., immediately before thermal cycling is begun, or by adding to the PCR reagents a single-stranded DNA binding protein which blocks reaction at temperatures below about 50 C. If the in situ PCR is performed on cellular preparations already attached to a microscope slide, thermal cycling also is facilitated by use of a thermal cycler sample block or compartment designed optimally to hold the microscope slide and any vapor barrier covering the slide.
Low-level malaria infections detected by a sensitive polymerase chain reaction assay and use of this technique in the...
The feasibility of using a sensitive polymerase chain reaction (PCR) to evaluate malaria vaccines in small group sizes was tested in 102 adult Gambian volunteers who received either the malaria vaccine regimen FP9 ME-TRAP/MVA ME-TRAP or rabies vaccine. All volunteers received the antimalarial drugs primaquine and Lapdap plus artesunate to eliminate malaria parasites. Volunteers in a further group received an additional single treatment with sulfadoxine-pyrimethamine (SP) to prevent new infections. There was substantially lower T-cell immunogenicity than in previous trials with this vaccine regimen and no protection against infection in the malaria vaccine group. Using the primary endpoint of 20 parasites per mL, no difference was found in the prevalence of low-level infections in volunteers who received SP compared with those who did not, indicating that SP did not reduce the incidence of very low-density infection. However, SP markedly reduced the incidence of higher density infections. These findings
Genomic Dna Extraction Protocol Phenol Chloroform - Online Lab Science Community
Lab Reagents Dna Extraction Laboratories manufactures the genomic dna extraction protocol phenol chloroform reagents distributed by Genprice. The Genomic Dna Extraction Protocol Phenol Chloroform reagent is RUO (Research Use Only) to test human serum or cell culture lab samples. To purchase these products, for the MSDS, Data Sheet, protocol, storage conditions/temperature or for the concentration, please contact DNA Extraction. Other Genomic products are available in stock. Specificity: Genomic Category: Dna Group: Extraction Protocol. Extraction Protocol information ...
A novel temperature control method for shortening thermal cycling time to achieve rapid polymerase chain reaction (PCR) in a...
TY - CONF. T1 - A novel temperature control method for shortening thermal cycling time to achieve rapid polymerase chain reaction (PCR) in a disposable polymer microfluidic device.. AU - Bu, Minqiang. AU - R. Perch-Nielsen, Ivan. AU - Sørensen, Karen Skotte. AU - Skov, Julia AU - Yi, Sun. AU - Bang, Dang Duong. AU - E. Pedersen, Michael AU - Hansen, Mikkel Fougt. AU - Wolff, Anders. PY - 2012. Y1 - 2012. N2 - We present a new temperature control method capable of effectively shortening the thermal cycling time of polymerase chain reaction (PCR) in a disposable polymer microfluidic device with external heater and temperature sensor. The method employs optimized temperature overshooting and undershooting steps to achieve a rapid ramping between the temperature steps for DNA denaturation, annealing and extension. The temperature dynamics within the microfluidic PCR chamber was characterized and the overshooting and undershooting parameters were optimized using the temperature dependent ...
Touchdown polymerase chain reaction - Wikipedia
The touchdown polymerase chain reaction or touchdown style polymerase chain reaction is a method of polymerase chain reaction by which primers avoid amplifying nonspecific sequences. The annealing temperature during a polymerase chain reaction determines the specificity of primer annealing. The melting point of the primer sets the upper limit on annealing temperature. At temperatures just above this point, only very specific base pairing between the primer and the template will occur. At lower temperatures, the primers bind less specifically. Nonspecific primer binding obscures polymerase chain reaction results, as the nonspecific sequences to which primers anneal in early steps of amplification will swamp out any specific sequences because of the exponential nature of polymerase amplification. The earliest steps of a touchdown polymerase chain reaction cycle have high annealing temperatures. The annealing temperature is decreased in increments for every subsequent set of cycles (the number of ...
Global Polymerase Chain Reaction Technologies Market Aanlysis, Growth, Demand, Study and Forecast 2019
 Global Polymerase Chain Reaction Technologies market study by product, technology and application. The study provides an in-depth analysis of the market current and future trends.
ReP USP - Detalhe do registro: A double-blinded, prospective study to define antigenemia and quantitative real time polymerase...
David-Neto E, Triboni AHK, Paula FJ, Vilas Boas LS, Machado CM, Agena F, Latif AZA, Alencar CS, Pierrotti LC, Nahas WC, Caiaffa-Filho HH, Pannuti CS. A double-blinded, prospective study to define antigenemia and quantitative real time polymerase chain reaction cutoffs to start preemptive therapy in low-risk, seropositive, renal transplanted recipients [Internet]. Clinical and Transplantation Research. 2014 ; 98( 10): 1077-1081.Available from: doi: 10.1097/TP. ...
Europe Real Time Polymerase Chain Reaction (qPCR) Market Analysis By Instrument (7500, Quantstudio Dx, ViiA 7 Dx, StepOne...
/PRNewswire/ -- Europe Real Time Polymerase Chain Reaction (qPCR) market is expected to reach over USD 395.8 million by 2022, according to a new report by...
Thermal cycler - Wikipedia
The earliest thermal cyclers were designed for use with the Klenow fragment of DNA polymerase I. Since this enzyme is destroyed during each heating step of the amplification process, new enzyme had to be added every cycle. This led to a cumbersome machine based on an automated pipettor, with open reaction tubes. Later, the PCR process was adapted to the use of thermostable DNA polymerase from Thermus aquaticus, which greatly simplified the design of the thermal cycler. While in some old machines the block is submerged in an oil bath to control temperature, in modern PCR machines a Peltier element is commonly used. Quality thermal cyclers often contain silver blocks to achieve fast temperature changes and uniform temperature throughout the block. Other cyclers have multiple blocks with high heat capacity, each of which is kept at a constant temperature, and the reaction tubes are moved between them by means of an automated process. Miniaturized thermal cyclers have been created in which the ...
Mutations of p53 gene can be detected in the plasma of patients with large bowel carcinoma. | Journal of Clinical Pathology
AIMS: To attempt to detect p53 gene mutations in the plasma of patients with large bowel carcinoma. METHODS: Plasma was collected from 20 control patients with no history of cancer and from 17 patients with large bowel carcinoma. Corresponding tumour and benign lymph node (control) samples for each of the carcinoma patients were obtained from paraffin blocks. A Dukes stage was determined for each tumour. DNA was extracted from the plasma samples and the paraffin embedded tissue using previously described methods. A nested primer polymerase chain reaction protocol was used for the amplification of exons 5 to 8 of the p53 gene. Cold single strand conformational polymorphism (SSCP) was performed on mini gels and silver stained. Abnormal bands were excised, the DNA eluted, and reamplified for automated dye termination sequencing. Any sample showing an apparent mutation was rechecked from the original extracted DNA sample at least three times. RESULTS: p53 gene mutations were not found in the ...
Potential bias of fungal 18S rDNA and internal transcribed spacer polymerase chain reaction primers for estimating fungal...
TY - JOUR. T1 - Potential bias of fungal 18S rDNA and internal transcribed spacer polymerase chain reaction primers for estimating fungal biodiversity in soil. AU - Anderson, Ian C. AU - Campbell, C. D.. AU - Prosser, James Ivor. PY - 2003. Y1 - 2003. N2 - Four fungal 18S rDNA and internal transcribed spacer (ITS) polymerase chain reaction (PCR) primer pairs were tested for their specificity towards target fungal DNA in soil DNA extracts, and their ability to assess the diversity of fungal communities in a natural grassland soil was compared. Amplified PCR products were cloned, and approximate to 50 clones from each library were sequenced. Phylogenetic analysis and database searches indicated that each of the sequenced cloned DNA fragments was of fungal origin for each primer pair, with the exception of the sequences generated using the 18S rDNA primers nu-SSU-0817 and nu-SSU-1196, where 35 of the 50 sequenced clones represented soil invertebrates. Although some of the primers have previously ...
Highly sensitive polymerase chain reaction methods show the frequent survival of residual recipient multipotent progenitors...
Abstract. Twenty-four male patients grafted for various pathologies with the marrow of a female donor and presenting a complete donor-type hematopoiesis when a
Reference genes for quantitative real-time polymerase chain reaction studies in soybean plants under hypoxic conditions
Quantitative real-time polymerase chain reaction (RT-qPCR) is a powerful tool used to measure gene expression. However, because of its high sensitivity, the method is strongly influenced by the quality and concentration of the template cDNA and by the amplification efficiency. Relative quantification is an effective strategy for correcting random and systematic errors by using the expression level of reference gene(s) to normalize the expression level of the genes of interest. To identify soybean reference genes for use in studies of flooding stress, we compared 5 candidate reference genes (CRGs) with the NormFinder and GeNorm programs to select the best internal control. The expression stability of the CRGs was evaluated in root tissues from soybean plants subjected to hypoxic conditions. Elongation factor 1-beta and actin-11 were identified as the most appropriate genes for RT-qPCR normalization by both the NormFinder and GeNorm analyses. The expression profiles of the genes for alcohol ...
Jadack, R.A., Yuenger, J., Ghanem, K.G., et al. (2006) Polymerase chain reaction detection of Y-chromosome sequences in vaginal...
Jadack, R.A., Yuenger, J., Ghanem, K.G., et al. (2006) Polymerase chain reaction detection of Y-chromosome sequences in vaginal fluid of women accessing a sexually transmitted disease clinic. Sexually Transmitted Diseases, 33, 22-25. doi10.1097/01.olq.0000194600.83825.81
Atomic force microscopic detection enabling multiplexed low-cycle-number quantitative polymerase chain reaction for biomarker...
TY - JOUR. T1 - Atomic force microscopic detection enabling multiplexed low-cycle-number quantitative polymerase chain reaction for biomarker assays. AU - Mikheikin, Andrey. AU - Olsen, Anita. AU - Leslie, Kevin. AU - Mishra, Bud. AU - Gimzewski, James K.. AU - Reed, Jason. PY - 2014/7/1. Y1 - 2014/7/1. N2 - Quantitative polymerase chain reaction is the current golden standard for quantification of nucleic acids; however, its utility is constrained by an inability to easily and reliably detect multiple targets in a single reaction. We have successfully overcome this problem with a novel combination of two widely used approaches: target-specific multiplex amplification with 15 cycles of polymerase chain reaction (PCR), followed by single-molecule detection of amplicons with atomic force microscopy (AFM). In test experiments comparing the relative expression of ten transcripts in two different human total RNA samples, we find good agreement between our single reaction, multiplexed PCR/AFM data, ...
Estimation of single-nucleotide polymorphism allele frequency by alternately binding probe competitive polymerase chain...
Fingerprint Dive into the research topics of Estimation of single-nucleotide polymorphism allele frequency by alternately binding probe competitive polymerase chain reaction. Together they form a unique fingerprint. ...
Quantification of the common deletion in human testicular mitochondrial DNA by competitive PCR assay using a chimaeric...
The common 4977 by deletion in mitochondrial DNA (Δ4977) is commonly used as an indicator of tissue deterioration in ageing and bioenergetic diseases. Deletion levels are normally measured by a serial dilution polymerase chain reaction (PCR) approach, where test reactions are compared with dilutions of control amplifications of DNA from a similar sized stable region of the mitochondrial genome. The end-point of this assay is the dilution that can just detect any PCR product; however, this is an inherently unstable measure. We constructed a chimaeric DNA construct that binds to both control and deletion primers with similar annealing properties. This was used in a competitive PCR assay to quantify Δ4977 in human testicular tissues that had been well-characterized using the serial dilution approach. We found the competitive assay to be highly replicable as it compares the PCR product of the construct with that of test DNA samples during the linear growth phase of the PCR reaction. Moreover, ...
Identification of Staphylococci by Polymerase Chain Reaction Directly from a Positive Blood Culture and Effect on Patient Care ...
Background: As one of the most common bloodstream infections worldwide, Staphylococcus aureus bacteremia places a major burden on health care. Implementation of a rapid, genetic-based diagnostic test may have important implications in the clinical management of patients with S. aureus bacteremia.. Objectives: The primary objective was to assess concordance between testing based on polymerase chain reaction (PCR) and the current gold standard, culture and sensitivity testing; the secondary objective was to assess the impact of this technology on patient care.. Methods: A pre-post intervention retrospective chart review was used to document the hospital course of patients with a diagnosis of S. aureus bacteremia before and after implementation of the PCR-based diagnostic system. Laboratory results from all patient samples subjected to PCR-based analysis following implementation of this system were compared with culture and sensitivity data for the same samples to determine accuracy of the new ...
Identification of Staphylococci by Polymerase Chain Reaction Directly from a Positive Blood Culture and Effect on Patient Care ...
Background: As one of the most common bloodstream infections worldwide, Staphylococcus aureus bacteremia places a major burden on health care. Implementation of a rapid, genetic-based diagnostic test may have important implications in the clinical management of patients with S. aureus bacteremia.. Objectives: The primary objective was to assess concordance between testing based on polymerase chain reaction (PCR) and the current gold standard, culture and sensitivity testing; the secondary objective was to assess the impact of this technology on patient care.. Methods: A pre-post intervention retrospective chart review was used to document the hospital course of patients with a diagnosis of S. aureus bacteremia before and after implementation of the PCR-based diagnostic system. Laboratory results from all patient samples subjected to PCR-based analysis following implementation of this system were compared with culture and sensitivity data for the same samples to determine accuracy of the new ...
Magnetic Beads Genomic DNA Extraction Kit (Blood) MB048/MB096 | Geneaid
The Magnetic Beads Genomic DNA Extraction Kit Blood was designed specifically for efficient genomic DNA purification from blood and buffy coat. DNA is bound to the surface of the magnetic beads and released using a proprietary buffer system.
Polymerase chain reaction
Understand how the DNA polymerase chain reaction works and is used in forensic science. Polymerase chain reaction (PCR) can be used in disease diagnosis, for example the diagnosis of avian influenza (bird flu)
Detection of human papillomavirus type 16 DNA sequences in archival cervical tissues by the polymerase chain reaction. -...
We have evaluated the polymerase chain reaction for the detection of viral DNA sequences in paraffin-embedded archival tissues. In 63 frozen cervical biopsy specimens that were taken from premalignant and invasive lesions, Southern blotting detected human papillomavirus (HPV) type 16 DNA in 28 (44%) of the samples. In the polymerase chain reaction analysis of the formalin-fixed, paraffin-embedded mirror biopsy specimens, 46 (73%) of the tissues were found to be positive for HPV type 16. In three Southern blotting-positive cases, the DNA of the paraffin-embedded sections was too scant or too degraded to allow the detection of HPV DNA by the polymerase chain reaction. In 21 Southern blotting-negative cases, HPV type 16 DNA could be demonstrated in the archival sections by the polymerase chain reaction technique--a sensitivity improvement of more than 80% over the standard method of HPV detection in tissues.
Journal of Current Medical Research and Practice - Prolactin level in patients with first-episode psychosis : Download PDF
This is a temporary file and hence do not link it from a website, instead link the URL of this page if you wish to link the PDF file ...
신개념 Genomic DNA 추출 키트- MagListoTM 5M Plant Genomic DNA Extraction Kit from Bioneer
MagListoTM 5M Plant Genomic DNA Extraction Kit 는 magnetic nano bead와 MagListoTM를 이용하여 Plant sample (leaf, root, seed) 에서 Genomic DNA를 빠르게 추출할 수 있는 획기적인 제품입니다. Magnetic nano bead와 자석을 이용해 세포 분쇄물 중 Genomic DNA만을 분리시키고 농축 및 정제하는 과정을 거치기 때문에 원심분리기를 사용하는 방법에 비해 빠르게 DNA를 분리 할 수 있습니다. 본 제품은 mini, midi, maxi scale의 prep을 위해 별도의 kit를 구매하지 않고 한 가지의 kit를 이용해 모두 prep 할 수 있으며 midi나 maxi prep을 위해 별도의 vacuum system이나 air pressure system을 구비 할 필요가 없는 것이 장점입니다.
Cy0 - A new method in Real-Time PCR quantification - Cy0 - About us ...
Cy0 is a new method in Real-time polymerase chain reaction quantification (PCR) analysis that does not require the assumption of equal efficiency between unknowns and standard curve.
Polymerase Chain Reaction (PCR) Products Market Size 2020 SWOT Analysis, Top Countries Data, Defination, Market Share, Growth...
The study of Polymerase Chain Reaction (PCR) Products market is a compilation of the market of Polymerase Chain Reaction (PCR) Products broken down into its entirety on the basis of types, application, trends and opportunities, mergers and acquisitions, drivers and restraints, and a global outreach. The detailed study also offers a board interpretation of the Polymerase Chain Reaction (PCR) Products industry from a variety of data points that are collected through reputable and verified sources. Furthermore, the study sheds a lights on a market interpretations on a global scale which is further distributed through distribution channels, generated incomes sources and a marginalized market space where most trade occurs.. Along with a generalized market study, the report also consists of the risks that are often neglected when it comes to the Polymerase Chain Reaction (PCR) Products industry in a comprehensive manner. The study is also divided in an analytical space where the forecast is predicted ...
Polymerase Chain Reaction Methods And Application
Polymerase chain reaction IPFS - polymerase chain reaction the many methods now used to rapid and widespread application as the polymerase chain reaction
Kontaktformular - SensoQuest GmbH
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Nasal swab samples and real-time polymerase chain reaction assays in community-based, longitudinal studies of respiratory...
Background Carefully conducted, community-based, longitudinal studies are required to gain further understanding of the nature and timing of respiratory viruses causing infections in the population. However, such studies pose unique challenges for field specimen collection, including as we have observed the appearance of mould in some nasal swab specimens. We therefore investigated the impact of sample collection quality and the presence of visible mould in samples upon respiratory virus detection by real-time polymerase chain reaction (PCR) assays. Methods Anterior nasal swab samples were collected from infants participating in an ongoing community-based, longitudinal, dynamic birth cohort study. The samples were first collected from each infant shortly after birth and weekly thereafter. They were then mailed to the laboratory where they were catalogued, stored at -80àand later screened by PCR for 17 respiratory viruses. The quality of specimen collection was assessed by screening for human ...
Rapid detection of CYP2C18 genotypes by real-time fluorescence polymerase chain reaction<...
TY - JOUR. T1 - Rapid detection of CYP2C18 genotypes by real-time fluorescence polymerase chain reaction. AU - Mizugaki, Michinao. AU - Hiratsuka, Masahiro. AU - Agatsuma, Yasuyuki. AU - Matsubara, Yoichi. AU - Fujii, Kunihiro. AU - Kure, Shigeo. AU - Narisawa, Kuniaki. PY - 2000/2. Y1 - 2000/2. N2 - In man, CYP2C19, a liver enzyme, plays an important role in the metabolism of several drugs. Mutation of the CYP2C19 gene results in a poor metaboliser phenotype. S-Mephenytoin hydroxylation genetic polymorphism is due to two mutations of the CYP2C19 gene, namely CYP2C19*2, located in exon 5, and CYP2C19*3, located in exon 4. CYP2C18 is also polymorphically expressed. The mutant alleles of this enzyme are CYP2C18ml, located in exon 2 and CYP2C18m2, located in the 5-flanking region. We have developed an allele-specific TaqMan polymerase chain reaction (PCR) assay with which to detect CYP2C18 mutant alleles. This assay combines hybridization of the TaqMan probe and allele-specific amplification ...
SARS-CoV-2 droplet digital polymerase chain reaction test
Bio-Rad provides a wide range of products for use in the support of COVID-19 diagnosis and confirmation, and offers a suite of molecular testing tools including Droplet Digital PCR (ddPCR) systems and the SARS-CoV-2 ddPCR test kit.. While real-time PCR provides an accessible, high-throughput option, the high sensitivity of ddPCR makes it well suited for screening samples in patients, providing the precision needed to resolve indeterminate test results ...
The Bio-Rad Droplet Digital PCR Publications Database
The introduction of digital PCR (dPCR) represented a paradigm shift in the field of quantitative polymerase chain reaction technology. Bio-Rad brought digital PCR to another level with the introduction of the revolutionary Droplet Digital PCR (ddPCR) technology. Today, with more than 3800 published studies and growing, nowhere is the power of Droplet Digital PCR technology further apparent than Bio-Rads expansive ddPCR Publications Database.
Monitoring NF-kappa B transactivation potential via real-time PCR quantification of I kappa B-alpha gene expression
The real-time PCR measure of I kappa B-alpha mRNA levels is a rapid, sensitive, and powerful method to quantify the transcriptional power of NF-kappa B. It can be easily used for clinical evaluation of NF-kappa B status.
Global Digital Polymerase Chain Reaction (dPCR) Market (Technology, Type, Application, End User and Geography) - Size, Share,...
Digital Polymerase Chain Reaction (dPCR) is an advancement of traditional polymerase chain reaction (PCR). The traditional PCR with its limited precision and accuracy often fail to amplify small samples of nucleic acid to a detectable level. This has evoked a need of better techniques to assess the minute quantities of DNA or RNA. dPCR is more sensitive and reliable technique with improved ability to quantify the absolute amount of nucleic acid. It divides the sample into large number of fragments, each containing either one or no template nucleic acid sequence. After DNA amplification, scoring is done with the help of fluorescence, counting the score as positive for the fraction containing template sequence and negative for the sample without the template sequence. The major factors driving the global digital polymerase chain reaction market are increasing demand for innovative diagnostic techniques, increasing disease awareness, need for early diagnosis of viral, infectious and genetic ...
Polymerase Chain Reaction (PCR) Market 2020 : Bio-Rad Laboratories, Abbott Laboratories, QIAGEN, Agilent Technologies, Inc.,...
This ready-to-refer market intelligence report on global Polymerase Chain Reaction (PCR) market entails a detailed analysis of the industrial ecosystem, followed by a highly reliable segment overview evaluated on multi-factor analysis, market size and dimensions in terms of volumetric gains and returns.. Get sample copy of Polymerase Chain Reaction (PCR) Market report @ https://www.orbispharmareports.com/sample-request/78970. Competitive Landscape Detailed Analysis: * Followed by constant and thorough research initiatives in data unraveling process pertaining to global Polymerase Chain Reaction (PCR) market, stringent curation processes have been directed to understand growth prognosis and development spanning across regional hubs and their respective performance and evaluation in terms of various macro and micro elements that decide further growth prognosis in global Polymerase Chain Reaction (PCR) market ...
High Pure PCR Template Preparation Kit from Roche Applied Science - a member of the Roche Group | SelectScience
Read independent reviews on High Pure PCR Template Preparation Kit from Roche Applied Science - a member of the Roche Group on SelectScience
OPUS 4 | Search
Background: Here we examined myocardial microRNA (miRNA) expression profile in a sensory neuropathy model with cardiac diastolic dysfunction and aimed to identify key mRNA molecular targets of the differentially expressed miRNAs that may contribute to cardiac dysfunction. Methods: Male Wistar rats were treated with vehicle or capsaicin for 3 days to induce systemic sensory neuropathy. Seven days later, diastolic dysfunction was detected by echocardiography, and miRNAs were isolated from the whole ventricles. Results: Out of 711 known miRNAs measured by miRNA microarray, the expression of 257 miRNAs was detected in the heart. As compared to vehicle-treated hearts, miR-344b, miR-466b, miR-98, let-7a, miR-1, miR-206, and miR-34b were downregulated, while miR-181a was upregulated as validated also by quantitative real time polymerase chain reaction (qRT-PCR). By an in silico network analysis, we identified common mRNA targets (insulin-like growth factor 1 (IGF-1), solute carrier family 2 facilitated ...
polymerase chain reaction steps
This provides single-stranded template for the next step temperature is remain about 94 -98°C. Repeated cycles of denaturation, primer annealling and extension carried out with the heat stable enzyme, Taq polymerase, leads to exponential increases in the target DNA sequences. And this is the sketch for the polymerase chain reaction. This process uses an enzyme derived from heat-resistant bacteria. 3.7. 1. heat to denature proteins (denaturation) ~98C 2. cool to anneal primers (short … By using this method you can amplify any region of gene which you want. Performing a Polymerase Chain Reaction 3. Scientist found T. aquaticus which lived in hot springs its DNA is most active at 70 degree thats way its DNA is most stable and become suitable enzyme for PCR used. Reverse transcription polymerase chain reaction (RT-PCR) is a laboratory technique combining reverse transcription of RNA into DNA (in this context called complementary DNA or cDNA) and amplification of specific DNA targets using ...
Performance of MiniPCR<sup>TM</sup> mini8, a portable thermal cycler |...
Performance of MiniPCRTM mini8, a portable thermal cycler - MiniPCRTM mini8 Thermal Cycler;GeneAmp®PCR system 9700;performance;STR multiplex kits;mtDNA HV1/HV2;
Browsing Research Output by Subject Real‐time polymerase chain reaction
Major depressive disorder (MDD) is associated with dysfunctional serotonergic and glutamatergic neurotransmission, and the genetic animal model of depression Flinders Sensitive Line (FSL) rats display alterations in these ...
Search of: 17659311 [PUBMED-IDS] - List Results - ClinicalTrials.gov
prevalence of falciparum malaria measured by qPCR (quantitative real time polymerase chain reaction), 12 months after the first administration of treatment with dihydroartemisinin-piperaquine and primaquine. (1017-13 and 23-15 ...
Used Bio-Rad T100 Thermal Cycler (202-9498) | PCR Thermal Cycler | Molecular Biology | BioSurplus
Price: $2699.00; Manufacturer: Bio-Rad; Item ID: 2029498; Warranty: 30-Day Money-Back Guarantee; Description: The Bio-Rad T100 is a compact thermal cycler perfect for laboratories with limited bench
PCR Thermal Cycler Testing Kits | Mandel Scientific
Is your Thermal Cycler heating and cooling properly? Achieving and maintaining the pre-programmed temperature? Giving uniform performance across the whole block? Gel Company offers Thermocycle Testing Kits which contain all the reagents needed for quick and easy performance testing on your Thermal Cycler. ...
TC-412 Thermal Cycler from Barloworld Scientific Ltd
TC-412 Thermal Cycler from Barloworld Scientific Ltd,The new TC-412 thermal cycler, flexible on your protocols, easy on your budget. ,The TC-412 is a high performance, high sample throughput instrument at a highly affordable price. ,,Key Features: , Flexible block format: The truly user-friendly fully interchangeable block sys,biological,biology supply,biology supplies,biology product
Esco | Spectrum 48 Real Time Thermal Cyclers
Discover Swift Spectrum 48 Thermal Cycler, an advanced real-time thermal cycler, which uses diode activation, PMT detection and a proprietary Peltier block design.
A multiplex polymerase chain reaction protocol for the simultaneous analysis of the glutathione S-transferase GSTM1 and GSTT1...
A multiplex polymerase chain reaction protocol for the simultaneous analysis of the glutathione S-transferase GSTM1 and GSTT1 polymorphisms
Use of 16S ribosomal DNA polymerase chain reaction to identify Haemophilus influenzae type B as the etiology of pericarditis in...
TY - JOUR. T1 - Use of 16S ribosomal DNA polymerase chain reaction to identify Haemophilus influenzae type B as the etiology of pericarditis in an infant . AU - Benson, Crystal. AU - Gantt, Soren. AU - Zerr, Danielle M.. AU - Qin, Xuan. AU - Abe, Patrick. N1 - Copyright: Copyright 2008 Elsevier B.V., All rights reserved.. PY - 2005/3. Y1 - 2005/3. UR - http://www.scopus.com/inward/record.url?scp=14944360668&partnerID=8YFLogxK. UR - http://www.scopus.com/inward/citedby.url?scp=14944360668&partnerID=8YFLogxK. U2 - 10.1097/01.inf.0000154593.22356.e2. DO - 10.1097/01.inf.0000154593.22356.e2. M3 - Letter. C2 - 15750478. AN - SCOPUS:14944360668. VL - 24. SP - 287. EP - 288. JO - Pediatric Infectious Disease Journal. JF - Pediatric Infectious Disease Journal. SN - 0891-3668. IS - 3. ER - ...
A Simple Polymerase Chain Reaction-Based Method for the Construction of Recombinase-Mediated Cassette Exchange Donor Vectors |...
For our initial test, we constructed the donor vector pBS-yin(B40XC), consisting of an intronless yellow gene flanked by inverted attB40 sites in the plasmid pBluescript (pBS). In constructing this donor, we subcloned the attB40 sites using complementary oligonucleotides with overhanging sticky ends that permitted ligation with restriction sites in pBS. We then used pBS-yin(B40XC) as a donor vector for RMCE via two methods. First, we co-injected the vector and mRNA encoding the ΦC31 integrase into embryos homozygous for an RMCE target in a yellow− white− background as described previously (Bateman et al. 2006). Among the progeny of surviving injectees, we were able to detect flies that had lost the white+ eye color produced by the mini-white gene in the target cassette and had gained yellow+ pigmentation, consistent with successful RMCE integration events. Although rates of integration by this method were relatively low (2-11%), they were consistent with control experiments using the ...
Get PDF - Development of a nested polymerase chain reaction test for the diagnosis of transmissible gastroenteritis of pigs
Rodríguez, E.; Betancourt, A.; Relova, D.; Lee, C.; Yoo, D.; Barrera, M., 2012: Development of a nested polymerase chain reaction test for the diagnosis of transmissible gastroenteritis of pigs
Quantitative real-time polymerase chain reaction (qRT-PCR) restriction fragment length polymorphism (RFLP) method for...
TY - JOUR. T1 - Quantitative real-time polymerase chain reaction (qRT-PCR) restriction fragment length polymorphism (RFLP) method for monitoring highly conserved transgene expression during gene therapy. AU - Bruzzone, Carol M.. AU - Belcher, John D.. AU - Schuld, Nathan J.. AU - Newman, Kristal A.. AU - Vineyard, Julie. AU - Nguyen, Julia. AU - Chen, Chunsheng. AU - Beckman, Joan D.. AU - Steer, Clifford J.. AU - Vercellotti, Gregory M.. PY - 2008/12. Y1 - 2008/12. N2 - Evaluation of the transfer efficiency of a rat heme oxygenase-1 (HO-1) transgene into mice requires differentiation of rat and mouse HO-1. However, rat and mouse HO-1 have 94% homology; antibodies and enzyme activity cannot adequately distinguish HO-1. We designed a quantitative real-time polymerase chain reaction (qRT-PCR) method to monitor HO-1 transcription relative to a housekeeping gene, GAPDH. The ratio of rat and mouse HO-1 mRNA could be estimated through restriction fragment length polymorphism (RFLP) analysis of the PCR ...
Repositorio da Producao Cientifica e Intelectual da Unicamp: Development of cycling probe-based real-time PCR system to detect...
In the present study, we developed a new real-time PCR system based on the cycling probe technology (CPT), which is composed of two single tube real-time PCR assays: the Fusarium genus-specific assay and the Fusanum solani species complex (FSSC)-specific assay with primers targeting the 28s ribosomal RNA gene. The Fusanum genus-specific assay was shown to be highly specific, detecting all reference Fusarium strains with no cross-reaction with other reference fungal strains, such as Aspergillus spp. and human DNA. The FSSC-specific assay also reacted very specifically with FSSC, except for a cross-reaction with Fusarium lunatum. To validate the real-time PCR system, we tested 87 clinical isolates of Fusarium spp. Identification results from the real-time PCR system were found to be 100% concordant with those from DNA sequencing of EF-1 alpha gene. The sensitivity testing also demonstrated high sensitivity, enabling detection of one copy of standard DNA with good reproducibility. Furthermore, both ...
Prevalence of and Risk Factors for Anal Oncogenic Human Papillomavirus Infection Among HIV-Infected Women in France in the...
Background. Little is known about the type-specific prevalence of anal human papillomavirus (HPV) infection and risk factors for anal high-risk (HR) HPV infection in human immunodeficiency virus (HIV)-infected women. Methods. A cross-sectional study of anal and cervical HPV infection was nested within a gynecological cohort of HIV-infected women. Specimens were tested for type-specific DNA using a polymerase chain reaction-based assay. Results. The study population consisted of 311 women with a median age of 45.3 years, of whom 42.8% originated from sub-Saharan Africa and 96.8% were receiving combination antiretroviral therapy. The median CD4 + cell count was 612/μL, and the HIV load was |50 copies/mL in 84.1%. HR-HPV types were detected in the anal canal in 148 women (47.6%) and in the cervix in 82 (26.4%). HPV-16 was the most prevalent type in both the anal canal (13.2% of women) and the cervix (5.1%). In multivariable analysis, factors associated with prevalent anal HR-HPV infection were CD4 + count
TPMT | Cancer Genetics Web
Thiopurine S-methyltransferase (TPMT) is an enzyme that converts thiopurine drugs into inactive metabolites. It is now well established that interindividual variation in sensitivity to thiopurines can be the result of the presence of genetic polymorphisms in the TPMT gene. The aim of this study was to determine the frequency and type of TPMT polymorphisms in the population of Serbia and Montenegro and to assess its relevance in the management of childhood acute lymphoblastic leukemia (ALL). Blood samples from 100 healthy adults and 100 children with ALL were analyzed for common mutations in the TPMT gene using polymerase chain reaction-based assays. The results revealed that allelic frequencies were 0.2% for TPMT*2, 3.2% for TPMT*3A, and 0.5% for TPMT*3B. A rare TPMT*3B allele was detected in 2 families. No TPMT*3C allele was found. The general pattern of TPMT-variant allele distribution as well as their frequencies in the population of Serbia and Montenegro, is similar to those determined for ...
Global preamplification simplifies targeted mRNA quantification | Scientific Reports
The need to perform gene expression profiling using next generation sequencing and quantitative real-time PCR (qPCR) on small sample sizes and single cells is rapidly expanding. However, to analyse few molecules, preamplification is required. Here, we studied global and target-specific preamplification using 96 optimised qPCR assays. To evaluate the preamplification strategies, we monitored the reactions in real-time using SYBR Green I detection chemistry followed by melting curve analysis. Next, we compared yield and reproducibility of global preamplification to that of target-specific preamplification by qPCR using the same amount of total RNA. Global preamplification generated 9.3-fold lower yield and 1.6-fold lower reproducibility than target-specific preamplification. However, the performance of global preamplification is sufficient for most downstream applications and offers several advantages over target-specific preamplification. To demonstrate the potential of global preamplification we
Differential Expression of the Multigene Family Encoding the Soybean Mitochondrial Alternative Oxidase | Plant Physiology
The alternative oxidase (AOX) of the soybean (Glycine max L.) inner mitochondrial membrane is encoded by a multigene family (Aox) with three known members. Here, the Aox2 and Aox3 primary translation products, deduced from cDNA analysis, were found to be 38.1 and 36.4 kD, respectively. Direct N-terminal sequencing of partially purified AOX from cotyledons demonstrates that the mature proteins are 31.8 and 31.6 kD, respectively, implying that processing occurs upon import of these proteins into the mitochondrion. Sequence comparisons show that the processing of plant AOX proteins occurs at a characteristic site and that the AOX2 and AOX3 proteins are more similar to one another than to other AOX proteins, including soybean AOX1. Transcript analysis using a polymerase chain reaction-based assay in conjunction with immunoblot experiments indicates that soybean Aox genes are differentially expressed in a tissue-dependent manner. Moreover, the relative abundance of both Aox2 transcripts and protein ...
BIO-410 Molecular Biology Techniques I (4.00)
BIO-410 Molecular Biology Techniques I (4.00). Introduces modern molecular biology techniques utilizing nucleic acids (DNA and RNA). Includes nucleic acid purification, quantitation, cloning and restriction enzyme digests. Advanced techniques include Southern and Northern analysis, polymerase chain reaction (PCR), real-time PCR and DNA sequencing. Stresses proficiency in techniques and proper analysis of results. Lab included. Credits: 4, Hours: (1/6/0/0), Arts & Sciences Elective Code: B. ...
Development and validation of real-time polymerase chain reaction assays specific to four species of Eimeria - RVC Research...
Blake, D P and Qin, Z and Cai, J and Smith, A L (2008) Development and validation of real-time polymerase chain reaction assays specific to four species of Eimeria. Avian Pathology, 37 (1). pp. 89-94. Full text not available from this repository ...
Species-specific polymerase chain reaction primers for simple detection of Bursaphelenchus fraudulentus (Nematoda:...
Affiliations: 1: Department of Biological Pest Control and Quarantine, Institute of Plant Protection, Wladyslawa Wegorka 20, Poznan, 60-318, Poland;, Email: [email protected]; 2: Department of Biological Pest Control and Quarantine, Institute of Plant Protection, Wladyslawa Wegorka 20, Poznan, 60-318, Poland, Department of Genetics, University of Valencia, Dr. Moliner 50, 46-100, Burjassot, Spain; 3: Department of Biological Pest Control and Quarantine, Institute of Plant Protection, Wladyslawa Wegorka 20, Poznan, 60-318, Poland ...
Genomic Changes of the 55 kDa Subunit of DNA Polymerase ε in Human Breast Cancer | Cancer Genomics & Proteomics
Background: DNA polymerases (Pols) represent potential candidates for cancer genes because of their central functions in DNA metabolism. Defects of some DNA Pols have shown cancer associations, but data on DNA polymerase (Pol) ε is limited. Materials and Methods: Twenty-four human breast cancer DNA samples and four control DNA samples were examined for possible mutation in the entire coding region of the 55 kDa small subunit of the human DNA Pol ε gene using polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) analysis of the DNA and sequence analysis. In addition, 20 control DNAs were studied with PCR-SSCP for the end of intron 18 and exon 19 region. Results: An AATT deletion was found at one location in intron 18 in 2 out of the 24 breast cancer cases (8%), but in none of the control cases. In addition, a single base transition was found in the cancer DNAs in intron 14, but the same changes were also found in the control DNAs, suggesting polymorphism. Conclusion: ...
A case of simultaneous multiple gastric cancers with p53 gene mutation.<...
TY - JOUR. T1 - A case of simultaneous multiple gastric cancers with p53 gene mutation.. AU - Lin, S. Y.. AU - Lee, M. D.. AU - Wang, C. K.. AU - Chen, Y. J.. AU - Chang, J. G.. PY - 1994/1. Y1 - 1994/1. N2 - Simultaneous multiple gastric cancers are rarely observed in clinical practice, and its association with p53 gene mutation has not been mentioned in any previous reports. We report a case of advanced gastric cancer with two primary lesions in the stomach who received total gastrectomy. Tumor and surrounding normal tissues from the surgical specimen were studied by using polymerase chain reaction-single strand conformation polymorphism analysis, restriction enzyme digestion method and direct sequencing. Point mutations of p53 gene at codon 248 (CGG--,TGG) were found in both primary tumor foci. The patient developed cancerous peritonitis eight months after the operation and expired six months later. This report suggests that p53 gene mutation can occur at an earlier stage in the tumorigenesis ...
Reagent Guide. Applied Biosystems StepOne and StepOnePlus Real-Time PCR Systems - PDF
Reagent Guide Applied Biosystems StepOne and StepOnePlus Real-Time PCR Systems Applied Biosystems StepOne and StepOnePlus Real-Time PCR Systems Reagent Guide Copyright 2008, 2010 Applied Biosystems. All
An accurate and rapid gender determination assay in single cells by the capillary polymerase chain reaction method<...
TY - JOUR. T1 - An accurate and rapid gender determination assay in single cells by the capillary polymerase chain reaction method. AU - Hashiba, Tsuyoshi. AU - Sueoka, Kou. AU - Kuroshima, Masako. AU - Asada, Hironori. AU - Kuji, Naoaki. AU - Yoshimura, Yasunori. N1 - Copyright: Copyright 2007 Elsevier B.V., All rights reserved.. PY - 1999. Y1 - 1999. N2 - Purpose: In preimplantation genetic diagnosis (PGD), a rapid and accurate assay has been required. We have therefore developed a capillary palymerase chain reaction (PCR) method using rapid thermal cycling programs to determine the gender of single amniocytes. Methods: Single amniocytes from each amniotic fluid sample were isolated by micromanipulation and their gender was determined by a multiplex PCR assay in a capillary tube, using primers that amplify a 308-bp DXZI and a 154-bp DYZI repeat sequence on the X and Y chromosomes, respectively. Results: All four thermal cycling programs, which took 180, 150, 120, and 90 min, were 100% accurate ...
Table 2 - Spoligotyping and Mycobacterium tuberculosis - Volume 11, Number 8-August 2005 - Emerging Infectious Disease journal ...
We evaluated the clinical usefulness of spoligotyping, a polymerase chain reaction-based method for simultaneous detection and typing of Mycobacterium tuberculosis strains, with acid-fast bacilli-positive slides from clinical specimens or mycobacterial cultures. Overall sensitivity and specificity were 97% and 95% for the detection of M. tuberculosis and 98% and 96% when used with clinical specimens. Laboratory turnaround time of spoligotyping was less than that for culture identification by a median of 20 days. In comparison with IS6110-based restriction fragment length polymorphism typing, spoligotyping overestimated the number of isolates with identical DNA fingerprints by ≈50%, but showed a 100% negative predictive value. Spoligotyping resulted in the modification of ongoing antimycobacterial treatment in 40 cases and appropriate therapy in the absence of cultures in 11 cases. The rapidity of this method in detection and typing could make it useful in the management of tuberculosis in a clinical
Welcome to CDC stacks | Spoligotyping and Mycobacterium tuberculosis - 14728 | Emerging Infectious Diseases
We evaluated the clinical usefulness of spoligotyping, a polymerase chain reaction-based method for simultaneous detection and typing of Mycobacterium tuberculosis strains, with acid-fast bacilli-positive slides from clinical specimens or mycobacterial cultures. Overall sensitivity and specificity were 97% and 95% for the detection of M. tuberculosis and 98% and 96% when used with clinical specimens. Laboratory turnaround time of spoligotyping was less than that for culture identification by a median of 20 days. In comparison with IS6110-based restriction fragment length polymorphism typing, spoligotyping overestimated the number of isolates with identical DNA fingerprints by approximately 50%, but showed a 100% negative predictive value. Spoligotyping resulted in the modification of ongoing antimycobacterial treatment in 40 cases and appropriate therapy in the absence of cultures in 11 cases. The rapidity of this method in detection and typing could make it useful in the management of ...
대량 sample을 위한 High-Throughput Genomic DNA Prep Kit- AccuPrep® Genomic DNA Extraction Kit for 96 well vacuum manifold
AccuPrep® Genomic DNA Extraction Kit for Biovac 96 Vacuum Manifold has been designed to quickly and conveniently extract genomic DNA from whole blood, buffy coat, lymphocytes, plasma, serum, body fluids, and cultured cells, simultaneously. The 96 samples can be handled without additional machinery such as a centrifuge. The genomic DNA is simply extracted with a vacuum pump and Biovac 96 Vacuum Manifold. This product is also available to other companies vacuum manifold system (QIAGEN, Promega and Axygen).
The American Journal of Tropical Medicine and Hygiene | Polymerase chain reaction-based identification and genotyping of...
A simple method for rapid identification of large numbers of Anopheles mosquitoes was developed based on polymerase chain reaction (PCR) amplification of the rDNA intergenic spacer and internal transcribed spacer 2. By means of previously described primers for the Anopheles gambiae and An. quadrimaculatus species complexes, rDNA was amplified simultaneously from 96 whole mosquitoes or parts. No homogenization or individual DNA preparation was necessary, and transfer of 96 samples to PCR reactions was performed simultaneously with a bacterial replicator. Control reactions indicate that the level of cross-contamination is negligible, and false-negative findings are rare. The method was tested on larvae, pupae, adult heads, whole adult males and females, and single tarsi. All parts except tarsi provided satisfactory template. Fresh, ethanol-preserved, dried, and frozen adults were also tested with similar results. The method was also tested for amplification of a single-copy gene, white. Results were
A Common LPA Null Allele Associates With Lower Lipoprotein(a) Levels and Coronary Artery Disease Risk | Arteriosclerosis,...
Approach and Results-The LPA null allele (rs41272114) was genotyped in the PROCARDIS case-control cohort (4073 CAD cases and 4225 controls). Lipoprotein(a) levels were measured in 909 CAD cases and 922 controls; apolipoprotein(a) isoform size was estimated using sodium dodecyl sulfate-agarose gel electrophoresis and a high-throughput quantitative polymerase chain reaction-based method. Null carriers are common (null allele frequency, 3%) and have significantly lower circulating lipoprotein(a) levels (P=2.1×10−10) and reduced CAD risk (odds ratio, 0.79 [0.66-0.97]; P=0.023) compared with noncarriers. An additive allelic model of apolipoprotein(a) isoform size, refined by null allele genotype and quantitative polymerase chain reaction values, showed a sigmoid relationship with lipoprotein(a) levels, with baseline levels for longer isoform alleles and progressively higher levels of lipoprotein(a) for shorter isoform alleles.. ...
Comparison of rapid methods of detection of cytomegalovirus in saliva with virus isolation in tissue culture. | Journal of...
Two rapid methods for the detection of cytomegalovirus (CMV) in saliva from congenitally and perinatally infected children were assessed by comparison with traditional virus isolation in tissue culture (TC). The polymerase chain reaction (PCR) was used to amplify a 300-bp segment of the CMV gB gene which was detected in ethidium bromide-stained agarose gels. A centrifugation-enhanced microtiter culture method with a monoclonal antibody for the detection of early-antigen fluorescent foci (DEAFF) was also used. Saliva specimens were collected with mouth swabs from children who were between the ages of 1 month and 14 years and who had either prenatal or perinatal CMV infection. One hundred sixty samples were tested by PCR and TC; 65 (40.6%) were found positive by TC, and 58 (36.8%) were found positive by PCR. Although four samples were found positive by PCR and negative by TC, saliva from seronegative and seropositive TC-negative adults were never found positive by PCR. One hundred fifty-two ...
Real-time quantitative PCR assay development and application for assessment of agricultural surface water and various fecal...
Aliarcobacter faecis and Aliarcobacter lanthieri are recently identified as emerging human and animal pathogens. In this paper, we demonstrate the development and optimization of two direct DNA-based quantitative real-time PCR assays using species-specific oligonucleotide primer pairs derived from rpoB and gyrA genes for A. faecis and A. lanthieri, respectively. Initially, the specificity of primers and amplicon size of each target reference strain was verified and confirmed by melt curve analysis. Standard curves were developed with a minimum quantification limit of 100 cells mL− 1 or g− 1 obtained using known quantities of spiked A. faecis and A. lanthieri reference strains in autoclaved agricultural surface water and dairy cow manure samples. Each species-specific qPCR assay was validated and applied to determine the rate of prevalence and quantify the total number of cells of each target species in natural surface waters of an agriculturally-dominant and non-agricultural reference watershed. In
Typing of mycobacteria using spoligotyping
Spoligotyping and Mycobacterium tuberculosis. Gori, Andrea; Bandera, Alessandra; Marchetti, Giulia; Esposti, Anna Degli; Catozzi, Lidia; Nardi, Gian Piero; Gazzola, Lidia; Ferrario, Giulio; Van Embden, Jan D. A.; Van Soolingen, Dick; Moroni, Mauro; Franzetti, Fabio // Emerging Infectious Diseases;Aug2005, Vol. 11 Issue 8, p1242 We evaluated the clinical usefulness of spoligotyping, a polymerase chain reaction-based method for simultaneous detection and typing of Mycobacterium tuberculosis strains, with acid-fast bacilli-positive slides from clinical specimens or mycobacterial cultures. Overall sensitivity and... ...
Evaluation of Microscopy and Polymerase Chain Reaction Methods for Identification of Trypanosoma Vivax in Cattle from Three...
Microscopy and Polymerase Chain Reaction technique was used to evaluate 150 blood samples from cattle in three abattoirs in Kaduna State to detect the presence of Trypanosoma vivax. Out of the 150 blood samples collected and tested for the presence of T. vivax using Microscopy,13 samples from Tudun-wada, Kawo and Makera abattoirs were found to positive giving a total prevalence of 26.0%. Tudun-wada abattoir had the highest prevalence with 16.0% while 3.0% and 2.0% for Kawo and Makera abattoirs respectively. Tudun- wada abattoir at 0.016 was seen to be significantly different at p. Key words: Abattoir, DNA, Microscopy, PCR, Primer, T. vivax. ...
A common LPA null allele associates with lower lipoprotein(a) levels and coronary artery disease risk. - Radcliffe Department...
OBJECTIVE: Increased levels of lipoprotein(a) are a highly heritable risk factor for coronary artery disease (CAD). The genetic determinants of lipoprotein(a) levels are mainly because of genetic variation in the apolipoprotein(a) gene (LPA). We have tested the association of a relatively common null allele of LPA with lipoprotein(a) levels and CAD risk in a large case-control cohort. We have also examined how null allele genotyping complements apolipoprotein(a) isoform typing to refine the relationship between LPA isoform size and circulating lipoprotein(a) levels. APPROACH AND RESULTS: The LPA null allele (rs41272114) was genotyped in the PROCARDIS (Precocious Coronary Artery Disease) case-control cohort (4073 CAD cases and 4225 controls). Lipoprotein(a) levels were measured in 909 CAD cases and 922 controls; apolipoprotein(a) isoform size was estimated using sodium dodecyl sulfate-agarose gel electrophoresis and a high-throughput quantitative polymerase chain reaction-based method. Null carriers are
SID.ir | DEVELOPMENT OF TWO MULTIPLEX POLYMERASE CHAIN REACTIONS FOR THE DETECTION OF ENTEROTOXIGENIC STRAINS OF STAPHYLOCOCCUS...
Download Free Full-Text of an article DEVELOPMENT OF TWO MULTIPLEX POLYMERASE CHAIN REACTIONS FOR THE DETECTION OF ENTEROTOXIGENIC STRAINS OF STAPHYLOCOCCUS AUREUS ISOLATED FROM FOODS
Evaluation of DNA extraction methods for the analysis of microbial community in biological activated carbon<...
TY - JOUR. T1 - Evaluation of DNA extraction methods for the analysis of microbial community in biological activated carbon. AU - Zheng, Lu. AU - Gao, Naiyun. AU - Deng, Yang. PY - 2012/2/1. Y1 - 2012/2/1. N2 - It is difficult to isolate DNA from biological activated carbon (BAC) samples used in water treatment plants, owing to the scarcity of microorganisms in BAC samples. The aim of this study was to identify DNA extraction methods suitable for a long-term, comprehensive ecological analysis of BAC microbial communities. To identify a procedure that can produce high molecular weight DNA, maximizes detectable diversity and is relatively free from contaminants, the microwave extraction method, the cetyltrimethylammonium bromide (CTAB) extraction method, a commercial DNA extraction kit, and the ultrasonic extraction method were used for the extraction of DNA from BAC samples. Spectrophotometry, agarose gel electrophoresis and polymerase chain reaction (PCR)-restriction fragment length ...
Article | Comparative evaluation of Polymerase Chain Reaction - Restriction Enzyme Analysis (PRA) and Sequencing of Heat shock...
Article: Pourahmad F, Adams A, Thompson K & Richards R (2009) Comparative evaluation of Polymerase Chain Reaction - Restriction Enzyme Analysis (PRA) and Sequencing of Heat shock protein 65 (hsp65) gene for identification of aquatic mycobacteria. Journal of Microbiological Methods, 76 (2), pp. 128-135. http://www.sciencedirect.com/science/journal/01677012; https://doi.org/10.1016/j.mimet.2008.09.021
A consensus on fungal polymerase chain reaction diagnosis? A United Kingdom-Ireland evaluation of polymerase chain reaction...
TY - JOUR. T1 - A consensus on fungal polymerase chain reaction diagnosis?. T2 - A United Kingdom-Ireland evaluation of polymerase chain reaction methods for detection of systemic fungal infections. AU - White, P. Lewis. AU - Barton, Richard. AU - Guiver, Malcolm. AU - Linton, Christopher J.. AU - Wilson, Steve. AU - Smith, Melvyn. AU - Gomez, Beatriz L.. AU - Carr, Michael J.. AU - Kimmitt, Patrick T.. AU - Seaton, Shila. AU - Rajakumar, Kumar. AU - Holyoake, Tessa. AU - Kibbler, Chris C.. AU - Johnson, Elizabeth. AU - Hobson, Richard P.. AU - Jones, Brian. AU - Barnes, Rosemary A.. N1 - Copyright: Copyright 2017 Elsevier B.V., All rights reserved.. PY - 2006/7. Y1 - 2006/7. N2 - The limitations of classical diagnostic methods for invasive fungal infections (IFIs) have led to the development of molecular techniques to aid in the detection of IFIs. Despite good published performance, interlaboratory reproduction of these assays is variable, and no consensus has been reached for an optimal ...
Environmental Engineering | AEESP
The Department of Civil and Environmental Engineering at Florida State University invites applications for a postdoctoral research associate position. The postdoc will work in a landfill leachate project. The ideal candidate should have a Ph.D. degree in Environmental Engineering or closely related fields. Candidates with previous experience in two or more of the following four areas are particularly encouraged to apply: 1) biological treatment of landfill leachate or wastewater; 2) geosynthetic clay liners; 3) quantitative real time polymerase chain reaction and next generation sequencing; 4) analytical techniques including SEM, TEM, XRD, Raman, IC, and TOC.. Review of applications will begin immediately, and will continue until the position is filled. The position is available immediately for a duration of one and a half year with the possibility of renewal based on satisfactory performance and funding. Interested applicants please contact [email protected] with a one-page cover letter, a CV, a ...
Can I use the Monarch® Genomic DNA Purification Kit to clean up gDNA that was extracted with phenol/chloroform? | NEB
Yes, the Monarch Genomic DNA Purification Kit can be used to clean up phenol/chloroform purified gDNA by following the protocol for Genomic DNA Cleanup. However, recovery may be lower than the usual 80% because phenol/chloroform purified DNA typically yields longer fragments. Although the use of preheated elution buffer in the Monarch protocol facilitates the elution of large gDNA fragments, the fraction of gDNA that is longer than 80 kb will be eluted less efficiently from the silica matrix ...
Integrin alpha subunit mRNAs are differentially expressed in early Xenopus embryos | Development
Adhesion of cells to extracellular matrix proteins is mediated, in large part, by transmembrane receptors of the integrin family. The identification of specific integrins expressed in early embryos is an important first step to understanding the roles of these receptors in developmental processes. We have used polymerase chain reaction methods and degenerate oligodeoxynucleotide primers to identify and clone Xenopus integrin alpha subunits from neurula-stage (stage 17) cDNA. Partial cDNAs encoding integrin subunits alpha 2, alpha 3, alpha 4, alpha 5, alpha 6 and an alpha IIb-related subunit were cloned and used to investigate integrin mRNA expression in early embryos by RNase protection assay and whole-mount in situ hybridization methods. Considerable integrin diversity is apparent early in development with integrins alpha 2, alpha 3, alpha 4, alpha 5 and alpha 6 each expressed by the end of gastrulation. Both alpha 3 and alpha 5 are expressed as maternal mRNAs. Zygotic expression of alpha 2, ...
Evaluation of the analytical and diagnostic performance of a digital droplet polymerase chain reaction (ddPCR) assay to detect...
TY - JOUR. T1 - Evaluation of the analytical and diagnostic performance of a digital droplet polymerase chain reaction (ddPCR) assay to detect Trypanosoma cruzi DNA in blood samples. AU - Ramírez, Juan David. AU - Herrera, Giovanny. AU - Hernández, Carolina. AU - Cruz-Saavedra, Lissa. AU - Muñoz, Marina. AU - Flórez, Carolina. AU - Butcher, Robert. PY - 2018/12/1. Y1 - 2018/12/1. N2 - Background: The recent development of novel Polymerase Chain Reaction (PCR) technologies that confer theoretical advantages over quantitative PCR has considerable potential in the diagnosis of low load infections, such as Trypanosoma cruzi in the chronic phase of Chagas disease. We evaluated the utility of the digital droplet (dd)PCR platform in the detection of T. cruzi infection. Methodology/Principal findings: We imported a validated qPCR assay targeting the T. cruzi satellite tandem repeat (TcSTR) region to the ddPCR platform. Following optimization, we tested and repeated a standard curve of TcI ...
Microbiology Society Journals | Rapid diagnosis of candidaemia by real-time PCR detection of Candida DNA in blood samples
This study prospectively evaluated an 18S rRNA gene-targeted real-time PCR approach in comparison with standard blood culture (BC) diagnostics for rapid diagnosis of candidaemia in a large study population of 384 patients, including 902 whole blood samples from 468 infectious episodes (IEs) of 329 adults and 55 children with haematological malignancies and various forms of immunodeficiency, and intensive care unit patients. Seven out of eight BC-proven cases (87.5 %) of candidaemia and seven out of twelve BC-positive samples (58.3 %) were positive by the Candida-specific PCR. A positive PCR result was also obtained for 28/460 BC-negative samples from IEs, including 8 patients with culture-confirmed Candida infection at primary sterile body sites. Of the PCR-positive, culture-negative patients, more than 50 % received systemic antifungal therapy. In 432/460 BC-negative IEs, the Candida specific-PCR was negative, resulting in a negative predictive value of 99.8 %. In conclusion, the Candida specific-PCR
Accuracy of genotyping of single-nucleotide polymorphisms by PCR-ELISA allele-specific oligonucleotide hybridization typing and...
Accuracy of genotyping of single-nucleotide polymorphisms by PCR-ELISA allele-specific oligonucleotide hybridization typing and by amplification refractory mutation ...
Real-time Polymerase Chain Reaction - The School of Biomedical Sciences Wiki
The Real-time Polymerase Chain Reaction is an improvised version of the original [[Polymerase Chain Reaction,Polymerase Chain Reaction]] (PCR,u,),/u, developed by Kary Mullis, who received the Nobel Prize in 1993 in Chemistry, and her coworkers during the mid-1980s. ,sup,(1),/sup ...
Evaluation of five DNA extraction methods in the detection of Salmonella enterica from meat using nested PCR
Polymerase Chain Reaction (PCR) based detection methods have received significant attention in food borne microbial pathogen detection. However, reliability and sensitivity of these methods are highly depending on the extraction of adequate amount of pure DNA using appropriate extraction method. Hence, selection of appropriate DNA extraction method is very important in PCR based detection of microbial pathogens. In this study, the extraction efficiency of five commonly used DNA extraction methods was evaluated. Salmonella enterica was used as experimental organism and five extraction methods were tested for their ability to extract DNA from spiked pork meat samples. Pork meat samples were incubated for four hours after being added a dilution series (100 - 103 CFU/mL) of Salmonella enterica culture. Then DNA was extracted from those samples by the five commonly used DNA extraction methods. Using extracted DNA, fliC gene of Salmonella was amplified by Nested PCR. Out of those five methods, the ...
Detection of hepatitis B virus in plasma using flow cytometric analyses of polymerase chain reaction-amplified DNA...
Blood donations are routinely screened by multiple serologic assays for antigens/antibodies associated with infection by blood-borne viruses, including hepatitis B virus (HBV), hepatitis C virus (HCV), human immunodeficiency viruses (HIV-1 and HIV-2), and human T-cell lymphotropic virus (HTLV-I and HTLV-II). A direct detection of these viruses would be more effective for the prevention of transfusion-transmitted infections than the indirect measurement of the variable host immune response to these agents. Because the polymerase chain reaction (PCR) for viral gene amplification offers the most sensitive and direct means of detecting viruses in blood, we have developed a nonisotopic PCR procedure for the detection of HBV, chosen as a prototype. The problems, common to previously described PCR methods, of nucleic acid extraction and inhibition of the PCR by plasma proteins were overcome by isolation of HBV from plasma by means of 450-microns polystyrene beads covalently coated with monoclonal antibody to
Real-time quantitative reverse transcriptase polymerase chain reaction.<...
TY - JOUR. T1 - Real-time quantitative reverse transcriptase polymerase chain reaction.. AU - Fan, Hongxin. AU - Robetorye, Ryan S.. PY - 2010. Y1 - 2010. N2 - The real-time quantitative reverse transcriptase polymerase chain reaction (RQ-PCR) has become the method of choice for the quantification of specific mRNAs. This method is fast, extremely sensitive, and accurate, requires only very small amounts of input RNA, and is relatively simple to perform. These characteristics have made it the method of choice for minimal residual disease monitoring such as in chronic myelogenous leukemia (CML). CML comprises approximately 20% of all leukemias and is characterized by a balanced (9;22) chromosomal translocation that results in the formation of a chimeric gene comprised of the BCR (breakpoint cluster region) gene and the ABL oncogene (BCR-ABL fusion gene). The chimeric gene encodes a fusion protein with constitutively increased tyrosine kinase activity, resulting in growth factor-independent ...
Direct solid-phase sequence analysis of the human p53 gene by use of multiplex polymerase chain reaction and alpha...
Among the candidate cancer-prognostic genes is the p53 tumor suppressor gene, which, when mutated, plays an important role in the development of many types of cancers. To facilitate robust large-scale DNA analysis of microdissected tumor biopsies, we describe a multiplex/nested PCR approach for a simultaneous outer amplification of exons 4-9 of the human p53 gene with parallel amplification of the HLA-DQB1 locus, involving a total of 14 primers. This approach reduces the required number of cells for analysis and avoids any variation in the amplifications of the individual p53 exons during the common outer amplification step. The HLA sequencing allows sample identification because the DQB1 locus is highly polymorphic and is thereby patient-specific. The p53 and HLA amplicons are analyzed by solid-phase sequencing in a semiautomated format. To improve the DNA sequence quality, we used 2-deoxyribonucleoside 5-O-1-thiotriphosphates in the sequencing reactions.. ...
Optimal conditions and specific characteristics of Vent exo- DNA polymerase in ligation-mediated polymerase chain reaction...
PubMed comprises more than 30 million citations for biomedical literature from MEDLINE, life science journals, and online books. Citations may include links to full-text content from PubMed Central and publisher web sites.
Background. Although high risk HPVs are associated with an increased risk of prostate cancer it is not known if they have a causal role. The purpose of this study is to investigate the potential role of human papilloma viruses (HPVs) in prostate cancer. The aims are (i) to investigate the presence and confirm the identity of high risk HPVs in benign prostate tissues prior to the development of HPV positive prostate cancer in the same patients, and (ii) to determine if HPVs are biologically active.. Methods. We used polymerase chain reaction (PCR) to identify HPVs in specimens from 52 Australian men with benign prostate biopsies who 1 to 10 years later developed prostate cancer. Immunohistochemistry (IHC) was used to assess the expression of HPV E7 oncoproteins, cytokeratin and prostate specific antigen (PSA).. We used RNASeq data from The Cancer Genome Atlas (TCGA) to identify possible HPV RNA sequences in prostate cancer.. Results. HPV screening using standard PCR was conducted on 28 of the 52 ...
High levels of HIV-1 in plasma during all stages of infection determined by competitive PCR | Science
Quantitative competitive polymerase chain reaction (QC-PCR) methods were used to quantify virion-associated human immunodeficiency virus type-1 (HIV-1) RNA in plasma from 66 patients with Centers for Disease Control stage I to IVC1 infection. HIV-1 RNA, ranging from 100 to nearly 22,000,000 copies per milliliter of plasma (corresponding to 50 to 11,000,000 virions per milliliter), was readily quantified in all subjects, was significantly associated with disease stage and CD4+ T cell counts, and decreased by as much as 235-fold with resolution of primary infection or institution of antiretroviral therapy. Plasma virus levels determined by QC-PCR correlated with, but exceeded by an average of 60,000-fold, virus titers measured by endpoint dilution culture. Quantitation of HIV-1 in plasma by QC-PCR may be useful in assessing the efficacy of antiretroviral agents, especially in early stage disease when conventional viral markers are often negative. ...
Mitochondrial haplotypebased identification of ethanolpreserved rootknot nematodes from Africa
The asexual root-knot nematodes (RKNs) (Meloidogyne spp.) exemplified by Meloidogyne incognita are widespread and damaging pests in tropical and subtropical regions worldwide. Comparison of amplification products of two adjacent polymorphic regions of the mitochondrial genome using DNA extracts of characterized RKN strains, including 15 different species, indicate that several species are derived from the same or closely related female lineages. Nevertheless, M. javanica, M. enterolobii, M. incognita, and other key species could each be assigned unique mitochondrial haplotypes based on polymerase chain reaction fragment size and restriction cleavage patterns. M. arenaria isolates did not group as a single haplotype, consistent with other reports of diversity within this species. To test the utility of this assay, we characterized ethanol-preserved samples from 103 single-species isolates from four countries in sub-Saharan Africa (Benin, Nigeria, Kenya, and Tanzania). Mitochondrial haplotypes ...
Absence of retroviral sequences in Graves disease<...
TY - JOUR. T1 - Absence of retroviral sequences in Graves disease. AU - Humphrey, M.. AU - Carr, F. E.. AU - Wartofsky, L.. AU - Djuh, Y. Y.. AU - Burman, K. D.. AU - Baker, J. R.. AU - Mosca, J.. AU - Drabick, J. J.. AU - Burke, D. S.. PY - 1991/1/5. Y1 - 1991/1/5. N2 - An earlier report of HIV-1 gene sequences in thyroid cell genomic DNA from patients with Graves disease prompted use of the polymerase chain reaction technique to identify such sequences in Graves disease thyroid tissue and in white blood-cells from these patients. We were unable to confirm the existence of HIV-1-related DNA sequences in Graves specimens.. AB - An earlier report of HIV-1 gene sequences in thyroid cell genomic DNA from patients with Graves disease prompted use of the polymerase chain reaction technique to identify such sequences in Graves disease thyroid tissue and in white blood-cells from these patients. We were unable to confirm the existence of HIV-1-related DNA sequences in Graves specimens.. UR - ...
Polymerase chain reaction. Polymerase chain reaction (PCR) assays are the most commonly used molecular technique to ... Sanger F, Nicklen S, Coulson AR (1977) DNA sequencing with chain-terminating inhibitors" Proceedings of the National Academy of ... This binding then sets off a chain of events that can be easily and definitively observed, depending on the test. More complex ... traditional PCR techniques require the use of gel electrophoresis to visualize amplified DNA molecules after the reaction has ...
Polymerase chain reaction. A polymerase chain reaction is a form of enzymatic DNA synthesis in the laboratory, using ... The term DNA synthesis can refer to DNA replication - DNA biosynthesis (in vivo DNA amplification), polymerase chain reaction ... cycles of repeated heating and cooling of the reaction for DNA melting and enzymatic replication of the DNA. ...
Polymerase chain reaction. Main article: Polymerase chain reaction. Polymerase chain reaction (PCR) is an extremely ... "Polymerase Chain Reaction (PCR) Fact Sheet". National Human Genome Research Institute (NHGRI). Retrieved 31 December 2016.. ... "Polymerase Chain Reaction (PCR)". National Center for Biotechnology Information. U.S. National Library of Medicine. Retrieved ... In this technique, DNA coding for a protein of interest is cloned using polymerase chain reaction (PCR), and/or restriction ...
Polymerase chain reaction (DNA). DNA profiling is today possible with even very small quantities of blood: this is commonly ...
Main articles: Taq Polymerase and History of polymerase chain reaction. In 1983, Mullis was working for Cetus Corporation as a ... Invention of polymerase chain reaction. Awards. William Allan Award (1990). Robert Koch Prize (1992). Nobel Prize in Chemistry ... K. Mullis, 1990, The unusual origin of the polymerase chain reaction. Scientific American, April 56-65. ... The Polymerase Chain Reaction, 1994, co-edited Francious Ferre and Richard A. Gibbs (Basel: Birkhauser) .mw-parser-output cite. ...
Polymerase Chain Reaction: An Alternative to Cloning *^ Brown TA (2002). "Section 2, Chapter 6: 6.1. The Methodology for DNA ... DNA can also be amplified using a procedure called the polymerase chain reaction (PCR). By using specific short sequences ... Kary Banks Mullis developed the polymerase chain reaction, providing a quick way to isolate and amplify a specific section of ... Each strand of DNA is a chain of nucleotides, matching each other in the center to form what look like rungs on a twisted ...
A polymerase chain reaction (PCR) test has been developed for the detection of Babesia from the peripheral blood. PCR may ... "Detection of Babesia microti by polymerase chain reaction". J. Clin. Microbiol. 30 (8): 2097-103. PMC 265450. PMID 1500517.. ...
Weier, HU; Gray, JW (Jul-Aug 1988). "A programmable system to perform the polymerase chain reaction". DNA (Mary Ann Liebert, ... is a laboratory apparatus most commonly used to amplify segments of DNA via the polymerase chain reaction (PCR). Thermal ... Later, the PCR process was adapted to the use of thermostable DNA polymerase from Thermus aquaticus, which greatly simplified ... The earliest thermal cyclers were designed for use with the Klenow fragment of DNA polymerase I. Since this enzyme is destroyed ...
Polymerase chain reactionEdit. Main article: Polymerase chain reaction. Polymerase chain reaction (PCR) is an extremely ... "Polymerase Chain Reaction (PCR) Fact Sheet". National Human Genome Research Institute (NHGRI). Retrieved 31 December 2016.. ... "Polymerase Chain Reaction (PCR)". www.ncbi.nlm.nih.gov. Retrieved 31 December 2016.. ... In this technique, DNA coding for a protein of interest is cloned using polymerase chain reaction (PCR), and/or restriction ...
Another method of screening is with polymerase chain reaction (PCR). Some libraries are stored as pools of clones and screening ...
Polymerase chain reaction(PCR). Additionally the development of recombinant DNA technology through the use of bacteria has ... From this microbe scientists isolated the enzyme Taq polymerase, which is now used in the powerful experimental technique, ... Fröls S, White MF, Schleper C (2009). "Reactions to UV damage in the model archaeon Sulfolobus solfataricus". Biochemical ... Terpe, Kay (1 November 2013). "Overview of thermostable DNA polymerases for classical PCR applications: from molecular and ...
Polymerase chain reaction is done by placing a mixture of the desired DNA, DNA polymerase, primers, and nucleotide bases into a ... "Polymerase chain reaction". "Gel electrophoresis". "Southern blot". White, Abraham (1959). "Principles of biochemistry". ... Molecular biology consists of different techniques including Polymerase chain reaction, Gel electrophoresis, and macromolecule ... Proteins are chains of amino acids that function, among other things, to contract skeletal muscle, as catalysts, as transport ...
For example, identification of novel DNA polymerases for polymerase chain reaction (PCR) reactions which synthesize DNA ... Garibyan, Lilit; Avashia, Nidhi (2013). "Polymerase Chain Reaction". Journal of Investigative Dermatology. 133 (3): 1-4. doi: ... With this in mind, 3173 Polymerase, another polymerase enzyme, now commonly used in RT-PCR reactions was discovered using the ... human DNA polymerase would denature during the denaturation step of the PCR reaction resulting in a non-functioning polymerase ...
Loop-mediated Isothermal Amplification
Alternative methods of polymerase chain reaction. In: Journal of pharmacy & bioallied sciences. Band 5, Nummer 4, Oktober 2013 ... Die LAMP verwendet eine strangversetzende DNA-Polymerase (z. B. Bst-DNA-Polymerase oder Bst 2.0) und läuft bei einer ... nicht-kovalente DNA-Katalysen und die hybridization chain reaction (HCR). Einzelnachweise[Bearbeiten , Quelltext bearbeiten ... K. Nagamine, T. Hase, T. Notomi: Accelerated reaction by loop-mediated isothermal amplification using loop primers. In: ...
by polymerase chain reaction". Croatian Medical Journal. 51 (4): 306-13. doi:10.3325/cmj.2010.51.306. PMC 2931435. PMID ... The inability to diagnose B. canis by SAT due to lack of cross-reaction is another drawback. False-negative SAT may be caused ... Demonstration of antibodies against the agent either with the classic Huddleson, Wright, and/or Bengal Rose reactions, either ... and demonstrate positive Bengal rose and Huddleston reactions. Gastrointestinal symptoms occur in 70% of cases and include ...
"Polymerase Chain Reaction (PCR)". www.ncbi.nlm.nih.gov. Retrieved 2020-05-16. "What is PCR?". Science Learning Hub. Retrieved ... Polymerase chain reaction, is the technology used for the purpose of copying particular DNA in a test tube. This method ...
The gold standard for diagnosis is PCR (polymerase chain reaction) helps DNA polymerase to create new strands of DNA equivalent ... "Polymerase Chain Reaction (PCR)". National Center for Biotechnology Information. Retrieved 2016-06-10. Pinart, Mariona; Rueda, ... but patients treated with paromomycin had a higher rate of adverse skin reactions. The treatments for other Leishmania species ...
Polymerase chain reaction is done by placing a mixture of the desired DNA, DNA polymerase, primers, and nucleotide bases into a ... Molecular biology consists of different techniques including Polymerase chain reaction, Gel electrophoresis, and macromolecule ... Proteins are chains of amino acids that function to contract skeletal muscle, function catalysts, transport molecules, and ... Proteins can facilitate biochemical processes, by lowering the activation energy of a reaction. Hemoglobins are also proteins, ...
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for the development of the polymerase chain reaction. 1992 格哈德·埃特爾. 德國 for the contributions to the new development of the ... for the discovery of Immunoglobulin E and mechanisms of IgE-mediated allergic reactions. ...
A polymerase chain reaction study". Arch Dermatol Res. 285 (5): 250-4. doi:10.1007/bf00371592. PMID 8397492. S2CID 22037398.CS1 ...
... is a variation of polymerase chain reaction (PCR) designed in 1988. The polymerase chain reaction (PCR) was ... "Polymerase Chain Reaction (PCR)". www.ncbi.nlm.nih.gov. Retrieved 2019-05-11. Riley J, Butler R, Ogilvie D, Finniear R, Jenner ... By doing this, the known sequence which is used to prime the PCR reaction at one side is introduced while the other is primed ...
Glossary of biology
polymerase chain reaction (PCR). A technique used in molecular biology to amplify a single copy or a few copies of a segment of ... endergonic reaction. Also called a nonspontaneous reaction or unfavorable reaction.. A type of chemical reaction in which the ... Also called the biosynthetic phase, light-independent reactions, dark reactions, or photosynthetic carbon reduction (PCR) cycle ... food chain. The chain of eating and getting nutrition which starts from a small herbivores animal and ends up at a big ...
... polymerase chain reaction) helps DNA polymerase to create new strands of DNA equivalent to template given. ... "Polymerase Chain Reaction (PCR)". National Center for Biotechnology Information. Retrieved 2016-06-10.. ... but patients treated with paromomycin had a higher rate of adverse skin reactions. ...
Tests that use polymerase chain reaction (PCR, aka nucleic acid amplification) to identify genes unique to N. gonorrhoeae are ... Traditionally, gonorrhea was diagnosed with Gram stain and culture; however, newer polymerase chain reaction (PCR)-based ... It is also possible for an individual to have an allergic reaction to the bacteria, in which case any appearing symptoms will ...
Alternatively, diagnosis and strain identification can be made using polymerase chain reaction (PCR). ...
Antibiotic sensitivity testing
Polymerase chain reaction is a method of identifying genes related to antibiotic susceptibility. In the PCR process, a ... Genetic testing, such as via polymerase chain reaction (PCR), DNA microarray, DNA chips, and loop-mediated isothermal ... and genetic methods such as polymerase chain reaction (PCR) testing have been available since the early 2000s. Research is ... Primers specific to a sought-after gene are added to a solution containing the DNA, and a DNA polymerase is added alongside a ...
Wilks (1989). "Two putative protein-tyrosine kinases identified by application of the polymerase chain reaction". PNAS. 86 (5 ...
doi:10.1006/jtbi.1996.0283 S. Schnell and C. Mendoza (1997). Theoretical description for polymerase chain reaction. Journal of ... Enzymological considerations for a theoretical description of the Quantitative Competitive Polymerase Chain Reaction (QC-PCR). ... His work has focused to resolve the ambiguities in the quantitative analysis and modeling of reactions inside cells. He has ... Schnell systematically investigated for the first time how the rate laws describing intracellular reactions vary as a function ...
Mullis, Kary B. (December 8, 1993). "Nobel Lecture - The Polymerase Chain Reaction". Fore J; Wiechers IR; Cook-Deegan R (2006 ... one of the most important enzymes in molecular biology because of its use in the polymerase chain reaction (PCR) DNA ... Kary Mullis and other investigators at Cetus Corporation discovered this enzyme could be used in the polymerase chain reaction ... and intellectual property on development and dissemination of the polymerase chain reaction: case study". J Biomed Discov ...
A primer dimer (PD) is a potential by-product in the polymerase chain reaction (PCR), a common biotechnological method. As its ... The primer design page of Leiden University Medical Center Patel, Ewing (2008). Polymerase Chain Reaction: Techniques and ... and these include initial inhibition of the DNA polymerase, or physical separation of reaction components reaction until the ... After an incubation of 1-5 minutes at 95 °C, the inhibitor is released and the reaction starts. Cold-sensitive Taq polymerase: ...
Ariosa v. Sequenom
The claim itself has two simple and conventional steps: first amplifying (by polymerase chain reaction, PCR) and then detecting ...
... detecting the viral RNA by polymerase chain reaction (PCR) and detecting proteins by enzyme-linked immunosorbent assay ( ... "This is the first time that all three countries - Guinea, Liberia and Sierra Leone - have stopped the original chains of ... The viral RNA polymerase, encoded by the L gene, partially uncoats the nucleocapsid and transcribes the genes into positive- ... reactions to the 1976 Ebola outbreak in Zaire. ...
This sequence is called a glycosylation sequon. The reaction catalyzed by OST is the central step in the N-linked glycosylation ... "Photocross-linking of nascent chains to the STT3 subunit of the oligosaccharyltransferase complex". J. Cell Biol. 161 (4): 715 ... Poly ADP ribose polymerase. *Sirtuin. Phosphoribosyltransferase. *Adenine phosphoribosyltransferase. *Hypoxanthine-guanine ... at the membrane of the endoplasmic reticulum and transferred to selected asparagine residues of nascent polypeptide chains by ...
... which is related to RNA polymerases found in bacteria. Chloroplasts also contain a mysterious second RNA polymerase that is ... Chloroplast polypeptide chains probably often travel through the two complexes at the same time, but the TIC complex can also ... its genes encode eleven subunits of a protein complex which mediates redox reactions to recycle electrons, which is similar ... The two RNA polymerases may recognize and bind to different kinds of promoters within the chloroplast genome. The ribosomes ...
Many modern molecular tests such as flow cytometry, polymerase chain reaction (PCR), immunohistochemistry, cytogenetics, gene ...
Polymerase chain reaction (PCR) tests for Lyme disease have also been developed to detect the genetic material (DNA) of the ... "Detection of Borrelia burgdorferi DNA by polymerase chain reaction in synovial fluid from patients with Lyme arthritis". The ... "Detection of Borrelia burgdorferi DNA by polymerase chain reaction in the urine and breast milk of patients with Lyme ... Chronic symptoms from an autoimmune reaction could explain why some symptoms persist even after the spirochetes have been ...
Pulmonya, ang malayang ensiklopedya
... gamit ang pag-culture o polymerase chain reaction(PCR), higit sa ibang mga pamamaraan. Ang nagdudulot na bagay ay natutukoy ...
Fluorescence in situ hybridization
Tagging can be done in various ways, such as nick translation, or Polymerase chain reaction using tagged nucleotides. ...
This can be detected at low cost using Polymerase Chain Reaction (PCR) of intron 1, followed by gel electrophoresis. Two bands ...
A direct confirmation can be obtained by reverse transcription polymerase chain reaction, where the genome of the virus is ... blood-sample testing with polymerase chain reaction is required.. A safe and effective vaccine against yellow fever exists, ...
Bioinformática, a enciclopedia libre
Polymerase Chain Reaction, reacción en cadea da polimerase) leva a poder facer a multiplicación de mostras de ADN, o que ... "A Short History of the Polymerase Chain Reaction". Methods in Molecular Biology 226.. ... "Markov Chain Monte Carlo" para análise bayesiano de problemas baseados en modelos probabilísticos. ... "DNA sequencing with chain-terminating inhibitors". Proceedings of National Academy of Sciences 74 (12). ...
... revolutionized molecular biology by allowing the polymerase chain reaction to be used in research as a simple and rapid ... This new appreciation of the importance and ubiquity of archaea came from using polymerase chain reaction (PCR) to detect ... Archaea exhibit a great variety of chemical reactions in their metabolism and use many sources of energy. These reactions are ... For example, thermostable DNA polymerases, such as the Pfu DNA polymerase from Pyrococcus furiosus, ...
Nuclear magnetic resonance spectroscopy of nucleic acids
... by performing polymerase chain reaction using dNTPs or NTPs derived from bacteria grown in an isotopically enriched environment ...
Reaction centers of photosynthetic bacteria: Feldafing-II-Meeting. 6. Berlin: Springer-Verlag. pp. 209-18. ISBN 3-540-53420-2. ... Ribosome-nascent chain complex (RNC). *Post-translational modification (functional groups · peptides · structural changes) ...
... long-chain enoyl-CoA hydratase, and long-chain thiolase. This deficiency can be classified into 3 main clinical phenotypes: ... "Vectorial proteomics reveal targeting, phosphorylation and specific fragmentation of polymerase I and transcript release ... long-chain-enoyl-CoA hydratase activity. Cellular component. • membrane. • mitochondrial outer membrane. • endoplasmic ... long-chain-3-hydroxyacyl-CoA dehydrogenase activity. • GO:0001948 protein binding. • catalytic activity. • transferase activity ...
ଜଳାତଙ୍କ - ଉଇକିପିଡ଼ିଆ
In: Polymerase Chain Reaction (PCR) for Human Viral Diagnosis Clewley JP, ed. Boca Raton: CRC. pp 125-145. ...
Reverse transcription polymerase chain reaction - විකිපීඩියා
Reverse transcription polymerase chain reaction (RT-PCR) යනු යම් නිශ්චිත වර්ගයක RNA ප්රමාණය මැනගැනීම සඳහා භාවිතාවන රසායනාගාර ... Reverse transcription polymerase chain reaction (RT-PCR). *Inverse polymerase chain reaction. *Nested polymerase chain reaction ... "https://si.wikipedia.org/w/index.php?title=Reverse_transcription_polymerase_chain_reaction&oldid=481677" වෙතින් සම්ප්රවේශනය ... Overlap extension polymerase chain reaction. *Multiplex polymerase chain reaction. *Multiplex ligation-dependent probe ...
α-1,4 glycogen chain)n + Pi ⇌ (α-1,4 glycogen chain)n-1 + α-D-glucose-1-phosphate. ... Although the reaction is reversible in solution, within the cell the enzyme only works in the forward direction as shown below ... The enzyme is specific to α1-4 chains, as the molecule contains a 30-angstrom-long crevice with the same radius as the helix ... Glycogen phosphorylase can act only on linear chains of glycogen (α1-4 glycosidic linkage). Its work will immediately come to a ...
ARNr 16S, a enciclopedia libre
"Phylogenetic analysis of Aquaspirillum magnetotacticum using polymerase chain reaction-amplified 16S rRNA-specific DNA". ...
Index of HIV/AIDS-related articles
... polymerase - Polymerase chain reaction (PCR) - polyneuritis - polypeptide - polyvalent vaccine - post-exposure prophylaxis (PEP ... adverse drug reaction - aerosolized - AETC - agammaglobulinemia - Agency for Healthcare Research and Quality (AHRQ) - AHRQ - ...
DNA paternity testing
The current techniques for paternity testing are using polymerase chain reaction (PCR) and restriction fragment length ... To satisfy the chain-of-custody legal requirements, all tested parties have to be properly identified and their specimens ... The DNA parentage test that follows strict chain of custody can generate legally admissible results that are used for child ... DNA test results are legally admissible if the collection and the processing follows a chain of custody. Similarly in Canada, ...
Հեպարին - Վիքիպեդիա՝ ազատ հանրագիտարան
Effects of heparin on polymerase chain reaction for blood white cells»։ J. Clin. Lab. Anal. 13 (3): 133-40։ 1999։ PMID 10323479 ... A Supply Chain Under Scrutiny։ Վերցված է նոյեմբերի 1, 2018 ...
... a first RNA version of the polymerase chain reaction (PCR).. Catalysis. The ability to catalyze simple chemical reactions- ... As each chain grew longer, it attracted more matching nucleotides faster, causing chains to now form faster than they were ... to describe this network of reactions. In November 2017, a team at the Scripps Research Institute identified reactions ... this allowed the team to isolate successful polymerases. The isolated RNA polymerases were again used for another round of ...
효소 - 위키백과, 우리 모두의 백과사전
Anfinsen CB (July 1973). "Principles that govern the folding of protein chains". 》Science》 181 (4096): 223-30. Bibcode:1973Sci ... Kopelman R (September 1988). "Fractal reaction kinetics". 》Science》 241 (4873): 1620-26. Bibcode:1988Sci...241.1620K. doi: ... DNA polymerase) 등이 그 예이다. 동일한 화학 반응을 촉해하는 서로 다른 효소들을 동질효소라고 한다. ... "Classification and Nomenclature of Enzymes by the Reactions they Catalyse". 》International Union of Biochemistry and Molecular ...
Tumor necrosis factor alpha
2007). "TNF Trafficking to Human Mast Cell Granules: Mature Chain-Dependent Endocytosis". The Journal of Immunology. 178 (9): ... negative regulation of transcription from RNA polymerase II promoter. • positive regulation of NF-kappaB transcription factor ... involved in systemic inflammation and is one of the cytokines that make up the acute phase reaction. It is produced chiefly by ... negative regulation of myosin-light-chain-phosphatase activity. • negative regulation of transcription, DNA-templated. • ...
RNA - Simple English Wikipedia, the free encyclopedia
The sequence of base pairs is transcribed from DNA by an enzyme called RNA polymerase. Then the mRNA moves from the nucleus to ... Transfer RNA (tRNA) is a short molecule of about 80 nucleotides which carries a specific amino acid to the polypeptide chain at ... This makes it the more suitable molecule to take part in cell reactions. ...
A type of polymerase chain reaction that detects the virus's RNA is more accurate. ... The most dangerous adverse effect is a severe allergic reaction to either the virus material itself or residues from the hen ... These core proteins and vRNA form a complex that is transported into the cell nucleus, where the RNA-dependent RNA polymerase ... Because of the absence of RNA proofreading enzymes, the RNA-dependent RNA polymerase that copies the viral genome makes an ...
Minimal residual disease
Generally this is achieved through the use of the polymerase chain reaction, a highly sensitive technique that underpins much ... Generally this is achieved through the use of reverse transcription of the RNA followed by polymerase chain reaction. RNA-based ...
Polymerase Chain Reaction
... ( PCR ) is a revolutionary molecular biology technique for enzymatically replicating DNA The … ... Polymerase Chain Reaction * 1. The Polymerase Chain Reaction (PCR) ,ul,,li,Polymerase chain reaction ( PCR ) is a revolutionary ... The extension temperature depends on the DNA-Polymerase ,/li,,/ul,Taq DNA Polymerase: This is a thermal stable enzyme isolated ... which provides a suitable chemical environment for the DNA-Polymerase ,/li,,/ul,,ul,,li,The PCR reaction is carried out in a ...
Polymerase Chain Reaction
... is covered by patents owned by Hoffmann-La Roche. DNA Amplification by Polymerase Chain Reaction. Our ... the polymerase chain reaction (PCR). The polymerase chain reaction was introduced to the scientific community at a conference ... Polymerase Chain Reaction. Mark V. Bloom, Ph.D.. DNA Learning Center,. Cold Spring Harbor Laboratory,. Cold Spring Harbor, New ... The polymerase chain reaction is a test tube system for DNA replication that allows a "target" DNA sequence to be selectively ...
PCR polymerase chain reaction
... A common method of amplifying target sequences of DNA in vitro by polymerase chain reaction (PCR ... You just viewed PCR polymerase chain reaction. Please take a moment to rate this material. ... Analysis of Food to Check for Genetic Modification by Polymerase Chain Reaction (PCR) ... and construction of the target sequence from free nucleotides by Taq polymerase. The cycle is repeated to demonstrate the power ...
Polymerase Chain Reaction - Xeroxing DNA
Polymerase Chain Reaction See Pioneer Profiles: Kary B. Mullis. See Carolina Tips: Polymerase Chain Reaction Return to About ... Polymerase Chain Reaction - Xeroxing DNA. National Center for Human Genome Research, National Institutes of Health. "New Tools ... The three parts of the polymerase chain reaction are carried out in the same vial, but at different temperatures. The first ... The three steps in the polymerase chain reaction - the separation of the strands, annealing the primer to the template, and the ...
History of Polymerase Chain Reaction (PCR)
The development of the polymerase chain reaction (PCR) has been a major breakthrough in the scientific world. Over time, the ... He added a second primer to the opposite strand and realized that repeated use of DNA polymerase will trigger a chain reaction ... The development of the polymerase chain reaction (PCR) has been a major breakthrough in the scientific world. Over time, the ... History of Polymerase Chain Reaction (PCR). News-Medical. https://www.news-medical.net/life-sciences/History-of-Polymerase- ...
Polymerase Chain Reaction (PCR) | NEB
Polymerase Chain Reaction (PCR). Return to DNA Amplification, PCR and qPCR The polymerase chain reaction (PCR) is a method to ... Home Applications DNA Amplification, PCR and qPCR Polymerase Chain Reaction (PCR) ... This overview will walk you through how the Polymerase Chain Reaction (PCR) works. ... DNA Replication with a Proofreading Polymerase Learn how proofreading polymerases recognize and correct mismatched bases. ...
CiteSeerX - Stochastic Modeling of Polymerase Chain Reaction and
2. The Polymerase Chain Reaction The PCR uses the mechanism of DNA replication. There are three steps in a PCR cycle. In the ... Introduction The polymerase chain reaction (PCR) is a method that uses test tubes in biological laboratories for producing ... The single-stranded sequences generated by denaturing are used as templates for the primers and the DNA polymerase. In the ... 2. The Polymerase Chain Reaction The PCR uses the mechanism of DNA replication. There are three steps in a PCR cycle. In the ...
PCR | PCR kits | Polymerase Chain Reaction
PCR, the polymerase chain reaction, is a core technique that has revolutionized molecular biology. In PCR, a DNA molecule is ... One of the most efficient methods for hot-start reactions can be achieved using antibodies to block Taq polymerase activity. ... Variations between thermostable DNA polymerases are used to optimize reactions for specific purposes. For example, in hot-start ... Taq Polymerase and Endpoint PCR. Includes high-performance GoTaq® DNA Polymerase, PCR buffers and master mixes for endpoint PCR ...
Polymerase Chain Reaction Primers | Physics Forums
In a PCR reaction DNA polymerase is responsible for building the new strand of DNA. However because of the mechanism by which ... Primers bind to the strand to give the polymerase something to extend. An old lecturer of mine used to give the analogy that ... DNA polymerase works it cannot just build a new strand opposite the old, it can only build off of existing DNA.. ...
Polymerase chain reaction - Wikipedia
The Polymerase Chain Reaction".. *^ "Determining Annealing Temperatures for Polymerase Chain Reaction". The American Biology ... In vitro Amplification of DNA by the Polymerase Chain Reaction *^ "Polymerase Chain Reaction (PCR)". National Center for ... "An Overview of Nanoparticle-Assisted Polymerase Chain Reaction Technology". An Overview of Nanoparticle‐Assisted Polymerase ... Main article: History of polymerase chain reaction. A 1971 paper in the Journal of Molecular Biology by Kjell Kleppe [no] and ...
Kary B. Mullis - Nobel Lecture: The Polymerase Chain Reaction - NobelPrize.org
So Im going to try to explain how it was that I invented the polymerase chain reaction. Theres a bit of it that will not ... The Polymerase Chain Reaction. In 1944 Erwin Schroedinger, stimulated intellectually by Max Delbrück, published a little book ... I opened a new file and named this one polymerase chain reaction. I didnt immediately try an experiment, but all summer I kept ... accomplishing the chain reaction. I was thinking of DNA:DNA interactions as being reversible with all the ramifications thereof ...
PCR - the polymerase chain reaction - Analytical Methods (RSC Publishing)
DNA-based procedures are becoming increasingly common within the analytical laboratory where the polymerase chain reaction (PCR ... PCR - the polymerase chain reaction Analytical Methods Committee, AMCTB No 59, Anal. Methods, 2014, 6, 333 DOI: 10.1039/ ... DNA-based procedures are becoming increasingly common within the analytical laboratory where the polymerase chain reaction (PCR ...
Patente US5538871 - In situ polymerase chain reaction - Google Patentes
... can be achieved by changing the way that the enzymatic reaction is started. Reaction initiation is delayed until the start of ... or by adding to the PCR reagents a single-stranded DNA binding protein which blocks reaction at temperatures below about 50 C. ... Improvements to the in situ polymerase chain reaction (PCR), a process of in vitro enzymatic amplification of specific nucleic ... The polymerase chain reaction (PCR) is a method for increasing by many orders of magnitude the concentration of a specific ...
Direct Automated Cycle Sequencing of Polymerase Chain Reaction Products | SpringerLink
The polymerase chain reaction (PCR) is well known for being a rapid and versatile method for the amplification of defined- ... The polymerase chain reaction (PCR) is well known for being a rapid and versatile method for the amplification of defined- ... Polymerase Chain Reaction Product Cycle Sequencing Microfuge Tube Single Nucleotide Substitution Guanine Cytosine These ... Mullis, K. B. and Faloona, F. A. (1987) Specific synthesis of DNA in vitro via a polymerase catalysed chain reaction. Methods ...
Polymerase chain reaction
... (PCR) can be used in disease diagnosis, for example the diagnosis of avian influenza (bird flu) ... Understand how the DNA polymerase chain reaction works and is used in forensic science. ... This technology exists - it is the polymerase chain reaction.. Like all brilliant ideas, the polymerase chain reaction is ... Polymerase chain reaction. PCR is a series of temperature-controlled reactions which enable us to amplify a very tiny sample of ...
Polymerase chain reaction
... (PCR) can be used in disease diagnosis, for example the diagnosis of avian influenza (bird flu) ... Understand how the DNA polymerase chain reaction works and is used in forensic science. ... Real time polymerase chain reaction. RT-PCR is a version of the polymerase chain reaction which allows the DNA to be amplified ... As the DNA polymerase moves along the template, the probe is cleaved (broken) between the reporter and quencher dye. This ...
Global Real-time PCR (Polymerase Chain Reaction) Systems
Polymerase Chain Reaction) Systems Market Opportunity Assessment to Reveal Lucrative Growth Prospects for Players - published ... Polymerase chain reaction (PCR) technology is used to amplify or make several copies of deoxyribonucleic acid (DNA) sequence of ... Polymerase chain reaction (PCR) technology is used to amplify or make several copies of deoxyribonucleic acid (DNA) sequence of ... Polymerase chain reaction (PCR) technology is used to amplify or make several copies of deoxyribonucleic acid (DNA) sequence of ...
Automation for Quantitative Polymerase Chain Reaction | Sigma-Aldrich
100 reactions. 400 reactions. S9194. SYBR Green JumpStart Taq ReadyMix for High-Throughput QPCR. 400 reactions. 2000 reactions ... 100 reactions. 500 reactions. S5193. SYBR Green JumpStart Taq ReadyMix without MgCl1. ... Quantitative PCR reaction setup is tedious and time consuming. Automation of reaction setup eliminates both human error and ... Automated methods have been developed and validated for Quantitative PCR reaction setup. The following resources are available ...
Detection of Minimal Residual Leukemia by Polymerase Chain Reactions | SpringerLink
Morgan GJ, Hughes T, Janssen JWG, Gow J, Guo AP, Goldman JM, Wiedemann LM, Bartram CR (1989) Polymerase chain reaction for ... In the following we will briefly summarize our recent experience with the application of polymerase chain reaction (PCR) ... Bone Marrow Transplantation Chronic Myeloid Leukemia Minimal Residual Disease Polymerase Chain Reaction Analysis Clinical ... Detection of Minimal Residual Leukemia by Polymerase Chain Reactions. In: Neth R., Frolova E., Gallo R.C., Greaves M.F., ...
Detection of Burkholderia pseudomallei in blood samples using polymerase chain reaction. - PubMed - NCBI
Quantitative polymerase chain reaction
... is a modification of the polymerase chain reaction used to rapidly measure ... Quantitative polymerase chain reaction Quantitative polymerase chain reaction (qPCR) ... Polymerase chain reaction techniques. Quantitative polymerase chain reaction (Q-PCR) , Real-time polymerase chain reaction (QRT ... Reverse transcription polymerase chain reaction (RT-PCR) , Inverse polymerase chain reaction , Nested polymerase chain reaction ...
07 | Real Time Polymerase Chain Reaction | Polymerase Chain Reaction
Use of the polymerase chain reaction for the specific and direct detection of Clostridium difficile in human feces. Rev. Infect ... Differentiation of bifidobacteria by use of pulsed-field gel electrophoresis and polymerase chain reaction. Int. J. Food ... Product differentiation by analysis of DNA melting curves during the polymerase chain reaction. Anal. Biochem. 245: 154-610. ... detection of Bifidobacterium strains in a pharmaceutical probiotic product and in human feces by polymerase chain reaction. ...
Biotechnical use of polymerase chain reaction for microbiological analysis of biological samples
... polymerase chain reaction (PCR) technology has been recognised as a rapid, sensitive and specific molecular diagnostic tool for ... Biotechnical use of polymerase chain reaction for microbiological analysis of biological samples Biotechnol Annu Rev. 2000;5:87 ... Since its introduction in the mid-80s, polymerase chain reaction (PCR) technology has been recognised as a rapid, sensitive and ... Polymerase Chain Reaction / methods* * Reverse Transcriptase Polymerase Chain Reaction / methods * Specimen Handling / methods ...
Gene splicing by overlap extension: tailor-made genes using the polymerase chain reaction. - PubMed - NCBI
... any pair of polymerase chain reaction products can be made to share a common sequence at one end. Under polymerase chain ... Gene splicing by overlap extension: tailor-made genes using the polymerase chain reaction.. Horton RM1, Cai ZL, Ho SN, Pease LR ... Extension of this overlap by DNA polymerase yields a recombinant molecule. This powerful and technically simple approach offers ... reaction conditions, the common sequence allows strands from two different fragments to hybridize to one another, forming an ...
2.01.03 Bluetongue | Agarose Gel Electrophoresis | Reverse Transcription Polymerase Chain Reaction
Reverse-transcription polymerase chain reaction (a prescribed test for international trade) Primer-directed amplification of ... Real-time reverse-transcription polymerase chain reaction tests Real-time RT-PCR is a sensitive method that can be used for the ... Reverse-transcription polymerase chain reaction RNA in ethanol is centrifuged at 12,000 g for 5 minutes. The ethanol is ... Reverse-transcription polymerase chain reaction (RT-PCR) technology has permitted rapid amplification of BTV and EHDV RNA in ...
Polymerase Chain Reaction (PCR) Market Forecast, Trend, Analysis & Compe
Global Polymerase Chain Reaction (PCR) Market - Scope of the Report The recent study by Fact.MR on the global polymerase chain ... reaction (PCR) market offers a 6-year forecast for the period of 2020-2026. The study analyzes ... Polymerase Chain Reaction (PCR) Market Forecast, Trend, Analysis & Competition Tracking - Global Market Insights 2020 to 2026. ... Global Polymerase Chain Reaction (PCR) Market: Size Evaluation. The global polymerase chain reaction (PCR) market has been ...
What the heck is the polymerase chain reaction? | Scienceline
What the heck is the polymerase chain reaction?. This molecular line dance is the most important process in genetic science. ... polymerase chain reaction.. Though the steps can seem complex, once you learn them, this molecular dance party becomes ... That repeated chain reaction leads to exponential growth in the number of identical DNA molecules available for testing. The ... allowing polymerase to grab free floating bases and rebuild the other side of the chain. ...
Polymerase chain reaction - Wikipedia
ISBN 978-0-879-69576-7. Chapter 8: In vitro Amplification of DNA by the Polymerase Chain Reaction "Polymerase Chain Reaction ( ... "DNA sequencing with Thermus aquaticus DNA polymerase and direct sequencing of polymerase chain reaction-amplified DNA". ... "An Overview of Nanoparticle-Assisted Polymerase Chain Reaction Technology". An Overview of Nanoparticle‐Assisted Polymerase ... Polymerase chain reaction (PCR) is a method widely used to rapidly make millions to billions of copies (complete copies or ...
Ultra-rapid flow-through polymerase chain reaction microfluidics using vapor pressure.
Polymerase chain reaction inhibitors - Wikipedia
PCR inhibitors are any factor which prevent the amplification of nucleic acids through the polymerase chain reaction (PCR). PCR ... or otherwise interfering with their interaction with the DNA polymerase, PCR is inhibited. In a multiplex PCR reaction, it is ... In order to try to assess the extent of inhibition that occurs in a reaction, a control can be performed by adding a known ... Of course, if any part of the inhibition occurring in the sample-derived reaction mixture is sequence-specific, then this ...
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- The cycles of amplification are described verbally as well as visually including the melting of DNA strands, the adherence of primers, and construction of the target sequence from free nucleotides by Taq polymerase. (merlot.org)
- This technique sought to simplify gene synthesis by using artificial primers and templates that help DNA polymerase to copy the desired gene segments. (news-medical.net)
- Meanwhile, the team used PCR for other applications by designing new primers and probes, which made the reaction more specific until the results were evident on agarose gel electrophoresis. (news-medical.net)
- During a typical PCR, template DNA (containing the region of interest) is mixed with deoxynucleotides (dNTPs), a DNA polymerase and primers. (neb.com)
- Primers are short segments of DNA that are complementary with the template DNA upstream of the region of interest and serve as recruitment sites for the polymerase. (neb.com)
- The single-stranded sequences generated by denaturing are used as templates for the primers and the DNA polymerase. (psu.edu)
- Primers bind to the strand to give the polymerase something to extend. (physicsforums.com)
- PCR employs two main reagents - primers (which are short single strand DNA fragments known as oligonucleotides that are a complementary sequence to the target DNA region) and a DNA polymerase . (wikipedia.org)
- A labelled probe is included in the PCR reagent mix in addition to the primers used in a traditional PCR reaction. (abpischools.org.uk)
- Two 22-base oligonucleotide primers, based on sequences from a specific DNA probe, were used for amplification of bacterial DNA by the polymerase chain reaction (PCR). (nih.gov)
- Both reactions are run for the same length of time in identical conditions (preferably using the same primers, or at least primers of similar annealing temperatures). (bionity.com)
- Using agarose gel electrophoresis separate the products of the reaction from their original DNA and spare primers. (bionity.com)
- PCR works the same way, but instead of a banquet hall full of wedding guests, it takes place in a test tube full of DNA templates and primers, polymerase, and nucleotides. (scienceline.org)
- Rychlik W (1993) Selection of primers for polymerase chain reaction. (els.net)
- Wenn sie erst einmal gebunden ist, fängt die Polymerase an freie Deoxy-Nukleotidtriphosphate, oder dNTPs, an das Ende des Primers, eines nach dem anderen von der 5' in die 3' Richtung anzufügen, um doppelsträngige DNA herzustellen. (jove.com)
- Sample DNA , nucleotides, DNA primers & thermostable DNA polymerase placed in PCR machine. (slideserve.com)
- PCR requires template DNA, Taq polymerase, di-deoxynucleic acids with each of the four DNA bases, Mg2+, primers and a buffer. (slideserve.com)
- We developed a single multiplex PCR reaction to detect ETEC, EPEC, EIEC, and STEC, using specific previously described ( 4 - 6 ) or new primers (GIBCO-BRL, Gaithersburg, MD) for diverse virulence traits ( Table 1 ). (cdc.gov)
- Taq DNA polymerase is known to be contaminated with low levels of bacterial DNA not originating from either Thermus aquaticus or Escherichia coli and is easily amplified using universal bacterial primers based on ribosomal gene sequences. (bmj.com)
- The specific Taq used in the study (AmpliTaq DNA polymerase) is well known for being unsuitable for bacterial PCR using pan-bacterial primers such that the company itself (Perkin-Elmer) has more recently introduced a "low DNA" Taq (Amplitaq LD) in order to reduce the size of the problem. (bmj.com)
- PCR is an enzymatic amplification process which requires DNA or RNA polymerase, primers and deoxynucleotide triphosphates in an appropriate buffer, and means of controlling the temperature during the various stages of the amplification process. (ubc.ca)
- During PCR, the template DNA is denatured to single-strands at a high temperature, then allowed to anneal with an excess of primers at a lower temperature, followed by synthesis of DNA or RNA from the primers at a temperature optimum for the specific polymerase enzyme. (ubc.ca)
- A sample of the supernatant containing chromosomal DNA is mixed with Taq DNA polymerase, oligonucleotide primers, the four deoxyribonucleotides, and the cofactor magnesium chloride. (ubc.ca)
- uses a thermostable polymerase (S-Tbr) that can extend from short primers ("smalligos") as short as 9 or 10 nucleotides. (primidi.com)
- Oligonucleotide consensus primers were designed to anneal to any of the four dengue virus types and amplify a 511-bp product in a reverse transcriptase-polymerase chain reaction (PCR). (asm.org)
- Using a patient's buffy coat and tick-bite site crust samples, we performed polymerase chain reaction (PCR) testing using Ehrlichia - or Anaplasma -specific primers. (ajtmh.org)
- The current study was designed to develop and validate a rapid and accurate diagnostic assay to detect two important genera of dermatophytes in dogs, Microsporum and Trichophyton differentially by a uniplex PCR reaction.18S ribosomal RNA gene of both the genera was used to design common forward and reverse primers. (thefreelibrary.com)
- The PCR process is completed in vitro and utilizes a DNA preparation containing the desired segment to be target sequence , two nucleotide primers about 20 bases long specific, the four deoxynucleoside triphosphates and a heat stable DNA polymerase. (assignmenthelp.net)
- Three essential steps to PCR (Figure 1) include (a) melting of the target (b) annealing of two oligonucleotide primers to the denatured DNA strands, and (c) primer extension by a thermostable DNA polymerase (123). (unl.edu)
- External primers were used to amplify a fragment of the expected size (441 bp) in all the samples evaluated using reverse transcriptase polymerase chain reaction (RT-PCR), but with very low intensity. (eurekamag.com)
- Specific detection of monkeypox virus by polymerase chain reaction. (nih.gov)
- Reactions were carried out on the ABI PRISM® 7700 Sequence Detection Systems. (sigmaaldrich.com)
- Detection of Burkholderia pseudomallei in blood samples using polymerase chain reaction. (nih.gov)
- Detection of bifidobacteria can be achieved using DNA extracted from human faeces as template in PCR reactions. (scribd.com)
- To obtain standardized quantitative results, external controls consisting of a DNA template for each target gene were used to generate linear standard curves over a 6 to 8 log range with detection of as few as 10 copies of amplicon per reaction. (bioone.org)
- Anamika Singh and Vijendra Kumar Kashyap, "Specific and Rapid Detection of Mycobacterium tuberculosis Complex in Clinical Samples by Polymerase Chain Reaction," Interdisciplinary Perspectives on Infectious Diseases , vol. 2012, Article ID 654694, 5 pages, 2012. (hindawi.com)
- Detection of specific polymerase chain reaction product by utilizing the 5'----3' exonuclease activity of Thermus aquaticus DNA polymerase. (pnas.org)
- The 5'----3' exonuclease activity of the thermostable enzyme Thermus aquaticus DNA polymerase may be employed in a polymerase chain reaction product detection system to generate a specific detectable signal concomitantly with amplification. (pnas.org)
- However, the increasing availability of viral diagnostic methods that use antigen detection or the polymerase chain reaction (PCR) allows the rapid identification of specific viral pathogens in febrile and afebrile infants. (aappublications.org)
- Nucleic acid hybridization techniques, exploited by colony hybridizations or polymerase chain reaction (PCR), apply a single detection method to a diversity of organisms. (cdc.gov)
- Rapid detection and typing of dengue viruses from clinical samples by using reverse transcriptase-polymerase chain reaction. (asm.org)
- Polymerase chain reaction (PCR) for the detection of food-borne pathogens. (ihs.com)
- Reverse-transcriptase-polymerase chain reaction (RT-PCR) has been applied successfully for detection of canine distemper virus (Agnihotri et al. (thefreelibrary.com)
- After dissection in search of flagellates in digestive tubes and identification of the species, female sandflies were submitted to the Multiplex Polymerase Chain Reaction (multiplex PCR) for detection of the fragment of the kDNA of Leishmania (Viannia) and the fragment from the IVS6 cacophony gene region of the phlebotomine insects. (scielo.br)
- Polymerase chain reaction (PCR) allows detection of Trypanosoma cruzi in blood throughout the course of Chagas' disease. (bmj.com)
- The Application Analysis of Multiplex Real-Time Polymerase Chain Reaction Assays for Detection of Pathogenic Bacterium in Peritoneal Dialysis-Associated Peritonitis. (bioportfolio.com)
- To estimate the clinical value of bacterial detection in peritoneal dialysis-associated peritonitis (PDAP) by multiplex real-time polymerase chain reaction (RT-PCR). (bioportfolio.com)
- Detection of cytomegalovirus (CMV) DNA by real-time polymerase chain reaction (rt-PCR) in dried blood spots (DBS) collected for newborn screening has been assessed for retrospective diagnosis of conge. (bioportfolio.com)
- Detection of viable but nonculturable Vibrio parahaemolyticus induced by prolonged cold-starvation using propidium monoazide real-time polymerase chain reaction. (bioportfolio.com)
- Detection of treponemal DNA in the CSF of patients with syphilis and HIV infection using the polymerase chain reaction. (bmj.com)
- The aim of this study was to develop a nested polymerase chain reaction (nested PCR) for the rapid detection of transmissible gastroenteritis virus (TGEV) of pigs. (eurekamag.com)
- Taq DNA Polymerase: This is a thermal stable enzyme isolated from thermophilic bacteria. (slideshare.net)
- Within a dividing cell, DNA replication involves a series of enzyme-mediated reactions, whose end result is a faithful copy of the entire genome. (accessexcellence.org)
- Within a test tube, PCR uses just one indispensable enzyme - DNA polymerase - to amplify a specific fraction of the genome. (accessexcellence.org)
- Thermal cycling exposes reactants to repeated cycles of heating and cooling to permit different temperature-dependent reactions-specifically, DNA melting and enzyme -driven DNA replication. (wikipedia.org)
- Almost all PCR applications employ a heat-stable DNA polymerase, such as Taq polymerase , an enzyme originally isolated from the thermophilic bacterium Thermus aquaticus . (wikipedia.org)
- Like other forms of polymerase chain reaction, the process is used to amplify DNA samples, via the temperature-mediated enzyme DNA polymerase . (bionity.com)
- DNA polymerase must be thermostable (37oC - 95oC used) to avoid fresh enzyme being added. (slideserve.com)
- The processivity, specificity, and fidelity of the polymerase enzyme used can largely determine the efficiency, reproducibility, and yield of the PCR reaction. (emdmillipore.com)
- KOD DNA Polymerase Is a Highly Efficient Proof Reading Enzyme. (emdmillipore.com)
- This enzyme provides the highest accuracy, yield and processivity of our proofreading DNA polymerases. (emdmillipore.com)
- replication also needs a primase enzyme to make primer for the polymerase (remember one of the rules for polymerases). (ubc.ca)
- And so, what this all boils down to: is the fact that you can get replication to work with only one enzyme - a workshorse DNA polymerase (i.e. (ubc.ca)
- 4. A heat stable DNA polymerase: This is an enzyme that is actually responsible for making a complementary DNA copy (i.e. replicating). (ubc.ca)
- We performed enzyme digestions after polymerase chain reaction amplification of internal transcribed spacer region and identified different restriction fragment length polymorphisms (RFLP) according to species and strains. (em-consulte.com)
- 96 QPCR reactions can be setup in less than 10 minutes. (sigmaaldrich.com)
- Quantitative polymerase chain reaction (qPCR) is a modification of the polymerase chain reaction used to rapidly measure the quantity of DNA , complementary DNA or ribonucleic acid present in a sample. (bionity.com)
- LONDON , July 12, 2016 /PRNewswire/ -- Europe Real Time Polymerase Chain Reaction (qPCR) market is expected to reach over USD 395.8 million by 2022, according to a new report by Grand View Research Inc. Technological advancements on the grounds of accuracy, portability, and cost-effectiveness are expected to serve this market as high-impact rendering drivers over the forecast period. (prnewswire.com)
- In real-time RT-PCR, also known as quantitative RT-PCR (RT-qPCR), data is collected while the reaction is occurring. (golden.com)
- Merck's One Step RT-PCR kits , NovaTaq™ Polymerase mastermixes and NovaQUANT™ qPCR assay kits make real-time PCR accessible, reproducible and standardized. (merckmillipore.com)
- DNA polymerases, whether from humans, bacteria, or viruses, cannot copy a chain of DNA without a short sequence of nucleotides to "prime" the process, or get it started. (accessexcellence.org)
- A PCR vial contains all the necessary components for DNA duplication: a piece of DNA, large quantities of the four nucleotides, large quantities of the primer sequence, and DNA polymerase. (accessexcellence.org)
- The Taq polymerase begins adding nucleotides to the primer and eventually makes a complementary copy of the template. (accessexcellence.org)
- The two DNA strands then become templates for DNA polymerase to enzymatically assemble a new DNA strand from free nucleotides , the building blocks of DNA. (wikipedia.org)
- Chemical compound made up of long chains of nucleotides joined together e.g. (abpischools.org.uk)
- Photochemical control of the polymerase chain reaction has been achieved through the incorporation of light-triggered nucleotides into DNA. (rsc.org)
- Direct solid-phase sequence analysis of the human p53 gene by use of multiplex polymerase chain reaction and alpha-thiotriphosphate nucleotides. (diva-portal.org)
Droplet Digital Polymera1
- We wished to develop an accurate quantitative protocol based on a droplet digital polymerase chain reaction (ddPCR) involving eight individual TaqMan™ reactions to detect simultaneously, without selective enrichment, Listeria spp. (frontiersin.org)
Ligation-mediated polymerase chain re1
- The most sensitive method uses ligation-mediated polymerase chain reaction (LM-PCR) to amplify all fragments of the genomic sequence ladder (3,4). (nih.gov)
- 3) Finally, the DNA-Polymerase has to fill in the missing strands. (slideshare.net)
- As the two strands separate, DNA polymerase makes a copy using each strand as a template. (accessexcellence.org)
- The three steps in the polymerase chain reaction - the separation of the strands, annealing the primer to the template, and the synthesis of new strands - take less than two minutes. (accessexcellence.org)
- Annealing of probe to one of the polymerase chain reaction product strands during the course of amplification generates a substrate suitable for exonuclease activity. (pnas.org)
- Denaturation is used to break DNA molecules into single strands, to permit the amplification reaction to proceed. (jove.com)
- It helps to use a DNA polymerase that is not denatured by the high temperature needed to separate the DNA strands. (biology-pages.info)
Agarose gel electrophor1
- With traditional PCR, the amount of product can only be worked out at the end of the reaction (the plateau phase), typically by agarose gel electrophoresis. (abpischools.org.uk)
Real-time quantitative polymera1
- Although real-time quantitative polymerase chain reaction is often marketed as RT-PCR, it should not to be confused with reverse transcription polymerase chain reaction , which is also referred to as RT-PCR, but is used to amplify RNA samples. (bionity.com)
- This study aimed to establish an accurate and sensitive polymerase chain reaction [PCR] technique for the diagnosis of active human brucellosis in Egypt. (who.int)
- The purpose of this study was to use the polymerase chain reaction (PCR) for diagnosis of enteroviral (EV) infection in febrile and afebrile infants ≤90 days of age to improve the understanding of the epidemiology of EV infection in this population. (aappublications.org)
- Comparative Evaluation of Immunochromatographic and Reverse Transcriptase Polymerase Chain Reaction based tests for Diagnosis of Canine Distemper. (thefreelibrary.com)
- 2014). This study is an attempt to compare polymerase chain reaction based molecular tests with commercially available IC based test for diagnosis of Canine distemper by testing rectal swabs and serum samples respectively from dogs suffering from gastroenteritis and suspected for Canine distemper. (thefreelibrary.com)
- Development and evaluation of a real-time polymerase chain reaction for fast diagnosis of sporotrichosis caused by Sporothrix globosa. (bioportfolio.com)
- polymerase chain reaction for fast diagnosis of sporotrichosis caused by Sporothrix globosa. (bioportfolio.com)
- The recent development of novel Polymerase Chain Reaction (PCR) technologies that confer theoretical advantages over quantitative PCR has considerable potential in the diagnosis of low load infections. (bioportfolio.com)
- Is Polymerase Chain Reaction in Neonatal Dried Blood Spots Reliable for the Diagnosis of Congenital Cytomegalovirus Infection? (bioportfolio.com)
- The polymerase chain reaction is a test tube system for DNA replication that allows a "target" DNA sequence to be selectively amplified, or enriched, several million-fold in just a few hours (Fig. 2). (accessexcellence.org)
- The first DNA polymerase was identified by Arthur Kornberg in 1957 during his studies on the DNA replication mechanism. (news-medical.net)
- 2. The Polymerase Chain Reaction The PCR uses the mechanism of DNA replication. (psu.edu)
- As PCR progresses, the DNA generated is itself used as a template for replication, setting in motion a chain reaction in which the original DNA template is exponentially amplified. (wikipedia.org)
- An optical instrument monitors PCR replication of DNA in a reaction apparatus having a temperature cycled block with vials of reaction ingredients including dye that fluoresces in presence of double-stranded DNA. (google.com)
- PCR involves replication of the DNA template by a thermostable DNA polymerase. (emdmillipore.com)
- this reaction is the same as that occurs in vivo during replication of the leading strand of a DNA duplex. (assignmenthelp.net)
- Analysis of major BCR-ABL1 mRNA by digital polymerase chain reaction is useful for prediction of international scale. (bioportfolio.com)
- New sensitive droplet digital PCR (polymerase chain reaction) assay found to be accurate in detecting hepatitis B virus (HBV) infection by measuring cccDNA in serum, single liver cells as well as processed tissue samples. (medindia.net)
- An oligonucleotide probe, nonextendable at the 3' end, labeled at the 5' end, and designed to hybridize within the target sequence, is introduced into the polymerase chain reaction assay. (pnas.org)
- A cocktail polymerase chain reaction assay to identify members of the Anopheles funestus (Diptera: Culicidae) group. (ajtmh.org)
- Evaluation of the analytical and diagnostic performance of a digital droplet polymerase chain reaction (ddPCR) assay to detect Trypanosoma cruzi DNA in blood samples. (bioportfolio.com)
Amplify a specific1
- He added a second primer to the opposite strand and realized that repeated use of DNA polymerase will trigger a chain reaction that will amplify a specific DNA segment, thus discovering the PCR technology. (news-medical.net)
- PCR inhibitors are any factor which prevent the amplification of nucleic acids through the polymerase chain reaction (PCR). (wikipedia.org)
- PCR inhibitors usually affect PCR through interaction with DNA or interference with the DNA polymerase. (wikipedia.org)
- By comparing the amplification of this template in the mixture to the amplification observed in a separate experiment in which the same template is used in the absence of inhibitors, the extent of inhibition in the investigated reaction mixture can be inferred. (wikipedia.org)
- As well as methods for the removal of inhibitors from samples before PCR, some DNA polymerases offer varying resistance to different inhibitors and increasing the concentration of the chosen DNA polymerase also confers some resistance to polymerase-targeted inhibitors. (wikipedia.org)
- Polymerase inhibitors present in both foods and enrichment media were removed efficiently. (biomedsearch.com)
- If the polymerase used was heat-susceptible, it would denature under the high temperatures of the denaturation step. (wikipedia.org)
- After synthesis, the cycle of denaturation, annealing and synthesis is repeated whereby the products of the previous amplification reaction serve as templates for the next round of amplification. (ubc.ca)
- It may be performed manually by heating the reaction components to the denaturation temperature (e.g., 95°C) before adding the polymerase. (primidi.com)
- First, we produced a cDNA copy of a portion of the viral genome in a reverse transcriptase reaction in the presence of primer D2 and then carried out a standard PCR (35 cycles of heat denaturation, annealing, and primer extension) with the addition of primer D1. (asm.org)
- The reaction mixture is first heated to a temperature between 90-98°C that ensures DNA denaturation and this step is called denaturation step. (assignmenthelp.net)
- To copy DNA, polymerase requires two other components: a supply of the four nucleotide bases and something called a primer. (accessexcellence.org)
- Once the primer is made, the polymerase can take over making the rest of the new chain. (accessexcellence.org)
- Although this technique used DNA polymerase repeatedly, similar to PCR, it employed only a single primer template complex, and exponential amplification was not possible using this technique. (news-medical.net)
- In the year 1977, Frederick Sanger identified a DNA sequencing method involving a DNA polymerase, a primer, and nucleotide precursors, for which he was he awarded the Nobel Prize in 1980. (news-medical.net)
- During the extension step (typically 68-72°C) the polymerase extends the primer to form a nascent DNA strand. (neb.com)
- Wenn die Primer an die DNA binden, starten sie die Reaktion, weil sie der Polymerase, einem Enzym, das die DNA repliziert, eine 3' Hydroxylgruppe zur Verfügung stellen. (jove.com)
- The third phase of a PCR cycle is "elongation" or "extension", when the DNA polymerase binds to the primer-template duplex and catalyzes synthesis of the product. (jove.com)
- First, a primer on the polymerase chain reaction (the lecture I would have liked to give in the lab if I had time). (ubc.ca)
- Because of the slow (arithmetic) amplification later in the reaction after the limiting primer has been used up, extra cycles of PCR are required. (primidi.com)
- A recent modification on this process, known as L inear- A fter- T he- E xponential-PCR (LATE-PCR), uses a limiting primer with a higher melting temperature (Tm) than the excess primer to maintain reaction efficiency as the limiting primer concentration decreases mid-reaction. (primidi.com)
- To order a free set of ABPI schools posters on the following topics: biotechnology, cloning, genetic engineering, unravelling the genome, polymerase chain reaction and stem cells, please fill in our order form . (abpischools.org.uk)
- MS2-like particles were produced through the synthesis of MS2 RNA genome by T7 RNA polymerase. (bireme.br)
- Therefore, before first round amplification, it is of paramount importance to pretreat the Taq polymerase with restriction enzymes (unpublished observations), and to include the first round negative control as a test sample in the nested PCR reaction. (bmj.com)
- The relationships between the parasitic nematodes of medical importance belonging to the genus Strongyloides was studied using a polymerase chain reaction (PCR)-linked restriction fragment length polymorphism approach. (ajtmh.org)
- This is a machine that heats and cools the reaction tubes within it to the precise temperature required for each step of the reaction. (slideshare.net)
- Since the Taq polymerase works best at around 75 degrees C (the temperature of the hot springs where the bacterium was discovered), the temperature of the vial is raised. (accessexcellence.org)
- Taking into account the time it takes to change the temperature of the reaction vial, 1 million copies can be ready in about three hours. (accessexcellence.org)
- Thermocyclers provide tight control over both the reaction temperature and the duration of each temperature step, ensuring efficient amplification. (neb.com)
- PCR is a series of temperature-controlled reactions which enable us to amplify a very tiny sample of DNA, producing enough material for it to be analysed or used in DNA profiling. (abpischools.org.uk)
- HOWEVER, we do have one problem: the high temperature used to open up the helical DNA molecule will basically knacker out any protein structures,including our polymerase. (ubc.ca)
- Hot-start/cold-finish PCR is achieved with new hybrid polymerases that are inactive at ambient temperature and are instantly activated at elongation temperature. (primidi.com)
- Temperature-controlled supply chains (cold chains) require an unbroken chain of refrigeration to maintain product quality and safety. (bioportfolio.com)
- Introduction The polymerase chain reaction (PCR) is a method that uses test tubes in biological laboratories for producing large amount of identical copies of a specific gene from small amount of complex molecules. (psu.edu)
- Successful polymerase chain reaction (PCR) is crucial for amplifying DNA sequences in order to study their function, either by sequencing, mutation, transcription, or expression of gene products. (emdmillipore.com)
- CG island methylation changes near the GSTP1 gene in prostatic carcinoma cells detected using the polymerase chain reaction: a new prostate cancer biomarker. (aacrjournals.org)
- Recently, simple and rapid assays for quantifying mRNA expression by real-time reverse transcription-polymerase chain reaction (RT-PCR) have been used for analysis of cytokine profiles in humans and other mammalian species. (bioone.org)
- Reverse transcription polymerase chain reaction, abbreviated RT-PCR, detects mRNA, pre-mRNA and noncoding RNAs. (golden.com)
- Major BCR-ABL1 mRNA in patients with chronic myeloid leukemia (CML) has generally been analysed by real-time polymerase chain reaction (PCR). (bioportfolio.com)
- A common method of amplifying target sequences of DNA in vitro by polymerase chain reaction (PCR) is described in this simplified yet accurate animation. (merlot.org)
- The polymerase chain reaction (PCR) is a method to rapidly amplify sequences of DNA. (neb.com)
- Improvements to the in situ polymerase chain reaction (PCR), a process of in vitro enzymatic amplification of specific nucleic acid sequences within the cells where they originate, can be achieved by changing the way that the enzymatic reaction is started. (google.es)
- The polymerase chain reaction (PCR) is well known for being a rapid and versatile method for the amplification of defined-target DNA sequences. (springer.com)
- In a multiplex PCR reaction, it is possible for the different sequences to suffer from different inhibition effects to different extents, leading to disparity in their relative amplifications. (wikipedia.org)
- Polymerase chain reaction (PCR) amplification of BLO nucleotide sequences was explored as a means to improve diagnostic techniques. (apsnet.org)
- The polymerase chain reaction, or PCR, is a powerful genetic technique that allows researchers to amplify DNA sequences of interest. (ubc.ca)
- The polymerase chain reaction (PCR) provides a simple, ingenious method to exponentially amplify specific DNA sequences by in vitro DNA synthesis. (unl.edu)
- Polymerase chain reaction ( PCR ) is a method widely used in molecular biology to make many copies of a specific DNA segment. (wikipedia.org)
- A mainstay of virtually every molecular biology lab, polymerase chain reaction (PCR) is an easy and affordable method for amplifying specific fragments of DNA. (sigmaaldrich.com)
- PCR (polymerase chain reaction) is an invaluable tool for molecular biology research. (thermofisher.com)
- The polymerase chain reaction, now widely used in research laboratories and doctor's offices, relies on the ability of DNA-copying enzymes to remain stable at high temperatures. (accessexcellence.org)
- When any cell divides, enzymes called polymerases make a copy of all the DNA in each chromosome. (accessexcellence.org)
- The DNA synthesis is carried out by DNA polymerase enzymes that are obtained from bacteria living in hot springs, such as Thermus aquaticus (Taq). (jove.com)
- KOD polymerase yields more product in fewer cycles compared to other PCR enzymes. (emdmillipore.com)
- Mullis, K. B. and Faloona, F. A. (1987) Specific synthesis of DNA in vitro via a polymerase catalysed chain reaction. (springer.com)
- an optical system adapted for being optically coupled to the one or more nucleic acid amplification reaction volumes accommodated by the support. (epo.org)
- b) an optical system including a detector operable to detect an optical signal related to the amount of amplified nucleic acid in the reaction mixture over a multiple-cycle period, without opening the reaction vessel once the amplification reaction is initiated. (epo.org)
- 2. The apparatus of claim 1, wherein the thermal cycler is capable of alternately heating and cooling a plurality of reaction vessels, each containing a said amplification reaction mixture. (epo.org)
- 7. The apparatus of any of the preceding claims, further comprising a reaction vessel adapted to contain a said amplification reaction mixture comprising a target DNA, reagents for said nucleic acid amplification, and a detectable nucleic acid binding agent. (epo.org)
- Ultimately this will entails the collection of a few components needed for a successful DNA amplification reaction. (ubc.ca)
- 9. A method in accordance with claim 1 wherein said first subset of PCR reagents consists of all PCR reagents except a nucleic acid polymerase. (google.es)
- Many scientists and other polymerase chain reaction (PCR) users believe samples need to be refrigerated immediately after thermal cycling finishes. (carolina.com)
- Since its introduction in the mid-80s, polymerase chain reaction (PCR) technology has been recognised as a rapid, sensitive and specific molecular diagnostic tool for the analysis of micro-organisms in clinical, environmental and food samples. (nih.gov)
- Reverse-transcription polymerase chain reaction (RT-PCR) technology has permitted rapid amplification of BTV and EHDV RNA in clinical samples, and RT-PCR-based procedures are now available. (scribd.com)
- The purpose of this study is to compare the diagnostic efficacy of nested and realtime polymerase chain reaction for Mycobacterium tuberculosis using EBUS-TBNA samples in patients with iso. (bioportfolio.com)
- To standardise and apply nested Polymerase Chain Reaction (n PCR) as a rapid and reliable laboratory diagnostic test to detect Toxoplasma gondii (T.gondii) in aqueous humor (AH) and peripheral blood samples of clinically suspected toxoplasma chorioretinitis patients. (arvojournals.org)
- Between May and October 2015 we analysed the serum samples of 140 patients with a suspicion of dengue, using ELISA and multiplex polymerase chain reaction. (who.int)
- Furthermore, automation has been facilitated, as reaction buffers and additional reagents can be rapidly changed without centrifugation or precipitation steps. (springer.com)
- Waiting for a reaction to finish can be inconvenient, though, and leaving a thermal cycler cooling overnight can severely reduce its useful life. (carolina.com)
- and monovalent cations, typically potassium (K) ions[better source needed] The reaction is commonly carried out in a volume of 10-200 μL in small reaction tubes (0.2-0.5 mL volumes) in a thermal cycler. (wikipedia.org)
- The thermal cycler heats and cools the reaction tubes to achieve the temperatures required at each step of the reaction (see below). (wikipedia.org)
- Q-PCR, Assembly PCR, Inverse PCR, RT-PCR and Digital PCR the Polymerase Chain Reaction (PCR) market to witness a value of US$12 billion by 2020. (reportsnreports.com)
- At initial stage of PCR , amplification of DNA segment is completed, two primary molecules can excess , the four deoxyribonucleosides triphosphates and the DNA polymerase are mixed together in the reaction mixture that has appropriate quantity of Mg2+. (assignmenthelp.net)
- TaqMan polymerase chain reaction was developed to quantify the number of Bifidobacterium . (ajol.info)
- Reverse-transcriptase polymerase chain reaction (RT-PCR) is a widely used method that enables transcriptional profiling and sequencing analysis on bulk populations of cells. (eurekamag.com)
- Lundeberg J., Pettersson B., Uhlén M. (2000) Direct DNA Sequencing of Polymerase Chain Reaction Products Using Magnetic Beads. (springer.com)
- To improve the DNA sequence quality, we used 2'-deoxyribonucleoside 5'-O-1-thiotriphosphates in the sequencing reactions. (diva-portal.org)
- 2-4 Although this level of contamination is insufficient to give a detectable amplification product after just one round of polymerase chain reaction (PCR), it is easily detected following nested amplification. (bmj.com)
- The levels of DNA contamination are easily detectable at the sensitivity (40 fg) for the second round PCR reported by this group of authors and neither in the text nor in the figures is there any mention of a first round negative control as a test sample in the nested PCR reaction. (bmj.com)
- It was determined whether Borrelia burgdorferi DNA may be detectable by polymerase chain reaction (PCR) in synovial membrane (SM) when PCR results from synovial fluid (SF) had become negative after antibiotic therapy. (bmj.com)
- Bone marrows of non-Hodgkin's lymphoma patients with a bcl-2 translocation can be purged of polymerase chain reaction-detectable lymphoma cells using monoclonal antibodies and immunomagnetic bead depletion. (harvard.edu)
- Amplification of bacterial 16S ribosomal DNA with polymerase chain reaction. (asm.org)
- Direct extraction of bacterial plasmids from food for polymerase chain reaction amplification. (biomedsearch.com)
- In this report we describe a simple and rapid technique using DNA affinity columns that permits direct extraction of bacterial plasmids from a variety of foods for polymerase chain reaction amplification. (biomedsearch.com)
- The contamination of Taq polymerase by bacterial DNA is now well established in the published press. (bmj.com)
- Conversely, real-time PCR (RT-PCR) allows for the simultaneous amplification and quantification of a targeted DNA molecule during a reaction. (abpischools.org.uk)
- The most sensitive quantification methods are done by the real-time polymerase chain reaction , where the amount of DNA is measured after each cycle of PCR by use of fluorescent markers. (bionity.com)
- A novel flow-through polymerase chain reaction (PCR) microfluidic system using vapor pressure was developed that can achieve ultra-rapid, small-volume DNA amplification on a chip. (biomedsearch.com)
- In this report, we describe a novel droplet-based microfluidic system for performing ~50000 single-cell RT-PCR reactions in a single experiment while consuming a minimal amount of reagent. (eurekamag.com)
- MilliporeSigma's molecular biologists work to develop the newest, best performing polymerases for customer use, optimizing conditions, buffer compositions, and cycling parameters to save you valuable time and resources. (emdmillipore.com)
- NovaTaq™ DNA polymerase is a premium quality recombinant form of Thermus aquaticus (Taq) DNA polymerase. (merckmillipore.com)
- NovaTaq™ master mixes have all the advantages of NovaTaq™ polymerase, plus the convenient setup and lower risk of cross contamination associated by the master mix format. (merckmillipore.com)
- NovaTaq™ Hot Start DNA Polymerase (red) shows more efficient real-time PCR amplification (left), yielding the same PCR product (right) compared to Supplier I Polymerase (green). (merckmillipore.com)
Limiting as the reaction progresses1
-  The amount of amplified product is determined by the available substrates in the reaction, which become limiting as the reaction progresses. (wikipedia.org)
- Kary Mullis had just conceived of a simple method for producing virtually unlimited copies of a specific DNA sequence in a test tube - the polymerase chain reaction (PCR) . (accessexcellence.org)
- Of course, if any part of the inhibition occurring in the sample-derived reaction mixture is sequence-specific, then this method will yield an underestimate of the inhibition as it applies to the investigate sequence(s). (wikipedia.org)
- In this study, we established a novel real-time polymerase chain reaction (PCR) method based on the internal transcribed spacer (ITS) sequence for the identification of S. globosa. (bioportfolio.com)
- The purpose of this study is to demonstrate the efficacy of active study vaccine in the prevention of reverse transcriptase polymerase chain reaction (RT-PCR) confirmed respiratory syncytial virus (RSV)-mediated lower respiratory tract disease (LRTD), when compared to placebo. (centerwatch.com)
- BACKGROUND Real-time reverse transcription polymerase chain reaction (RT-PCR) is routinely used to detect viral infections. (bireme.br)
- An analysis of the transcript level of the 10 members of the family has been performed using the technique of real-time quantitative reverse transcriptase-polymerase chain reaction. (plantphysiol.org)
- Polymerase Chain reaction (PCR) was developed to amplify and study genomic DNA (gDNA). (coursehero.com)
- DNA-based procedures are becoming increasingly common within the analytical laboratory where the polymerase chain reaction (PCR) has become an indispensable technique. (rsc.org)
- If the template contains a G, it adds a C to the new chain, and so on to the end of the DNA strand. (accessexcellence.org)
- In a PCR reaction DNA polymerase is responsible for building the new strand of DNA. (physicsforums.com)
- However because of the mechanism by which DNA polymerase works it cannot just build a new strand opposite the old, it can only build off of existing DNA. (physicsforums.com)
- Each ReadyMix contains Sigma's antibody mediated hot start mechanism, JumpStart™ Taq DNA Polymerase, for highly specific amplification. (sigmaaldrich.com)
- In order to try to assess the extent of inhibition that occurs in a reaction, a control can be performed by adding a known amount of a template to the investigated reaction mixture (based on the sample under analysis). (wikipedia.org)
- KOD-XL DNA polymerase amplification results in a mixture of blunt and 3'-dA products while other KOD DNA polymerases generate blunt end products. (emdmillipore.com)
- In the case of RT-PCR, these are labelled with a fluorescent reporter dye at the 5' end and a quencher dye at the 3' end and only produce fluorescence when DNA polymerase moves along the template and cleaves (splits) the probe. (abpischools.org.uk)
- As the DNA polymerase moves along the template, the probe is cleaved (broken) between the reporter and quencher dye. (abpischools.org.uk)
- During amplification, the 5'----3' exonuclease activity of T. aquaticus DNA polymerase degrades the probe into smaller fragments that can be differentiated from undegraded probe. (pnas.org)
- Some investigators recognize the occurrence of OHL under the subclinical form, which is identified by the cytophatics effects of EBV by histopathology or cytopathology, and DNA viral confirmation by polymerase chain reaction (PCR) (Fraga-Fernandez et al. (scielo.br)
- Polymerase chain reaction (PCR) is a method widely used to rapidly make millions to billions of copies (complete copies or partial copies) of a specific DNA sample, allowing scientists to take a very small sample of DNA and amplify it (or a part of it) to a large enough amount to study in detail. (wikipedia.org)
- The DNA polymerase copies the DNA again. (slideserve.com)