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.
A variation of the PCR technique in which cDNA is made from RNA via reverse transcription. The resultant cDNA is then amplified using standard PCR protocols.
The sequence of PURINES and PYRIMIDINES in nucleic acids and polynucleotides. It is also called nucleotide sequence.
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-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.
Short sequences (generally about 10 base pairs) of DNA that are complementary to sequences of messenger RNA and allow reverse transcriptases to start copying the adjacent sequences of mRNA. Primers are used extensively in genetic and molecular biology techniques.
Methods used for detecting the amplified DNA products from the polymerase chain reaction as they accumulate instead of at the end of the reaction.
A DNA-dependent RNA polymerase present in bacterial, plant, and animal cells. It functions in the nucleoplasmic structure and transcribes DNA into RNA. It has different requirements for cations and salt than RNA polymerase I and is strongly inhibited by alpha-amanitin. EC 2.7.7.6.
Binary classification measures to assess test results. Sensitivity or recall rate is the proportion of true positives. Specificity is the probability of correctly determining the absence of a condition. (From Last, Dictionary of Epidemiology, 2d ed)
Deoxyribonucleic acid that makes up the genetic material of viruses.
The order of amino acids as they occur in a polypeptide chain. This is referred to as the primary structure of proteins. It is of fundamental importance in determining PROTEIN CONFORMATION.
Deoxyribonucleic acid that makes up the genetic material of bacteria.
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).
The insertion of recombinant DNA molecules from prokaryotic and/or eukaryotic sources into a replicating vehicle, such as a plasmid or virus vector, and the introduction of the resultant hybrid molecules into recipient cells without altering the viability of those cells.
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).
The biosynthesis of RNA carried out on a template of DNA. The biosynthesis of DNA from an RNA template is called REVERSE TRANSCRIPTION.
The genetic constitution of the individual, comprising the ALLELES present at each GENETIC LOCUS.
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 2.7.7.7.
A method (first developed by E.M. Southern) for detection of DNA that has been electrophoretically separated and immobilized by blotting on nitrocellulose or other type of paper or nylon membrane followed by hybridization with labeled NUCLEIC ACID PROBES.
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.
Any detectable and heritable change in the genetic material that causes a change in the GENOTYPE and which is transmitted to daughter cells and to succeeding generations.
The phenotypic manifestation of a gene or genes by the processes of GENETIC TRANSCRIPTION and GENETIC TRANSLATION.
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 2.7.7.7.
Methods for using more than one primer set in a polymerase chain reaction to amplify more than one segment of the target DNA sequence in a single reaction.
Variation occurring within a species in the presence or length of DNA fragment generated by a specific endonuclease at a specific site in the genome. Such variations are generated by mutations that create or abolish recognition sites for these enzymes or change the length of the fragment.
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 2.7.7.7.
A multistage process that includes cloning, physical mapping, subcloning, determination of the DNA SEQUENCE, and information analysis.
Deoxyribonucleic acid that makes up the genetic material of protozoa.
Electrophoresis in which agar or agarose gel is used as the diffusion medium.
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.
Single-stranded complementary DNA synthesized from an RNA template by the action of RNA-dependent DNA polymerase. cDNA (i.e., complementary DNA, not circular DNA, not C-DNA) is used in a variety of molecular cloning experiments as well as serving as a specific hybridization probe.
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.
Variant forms of the same gene, occupying the same locus on homologous CHROMOSOMES, and governing the variants in production of the same gene product.
Ribonucleic acid that makes up the genetic material of viruses.
Macromolecular molds for the synthesis of complementary macromolecules, as in DNA REPLICATION; GENETIC TRANSCRIPTION of DNA to RNA, and GENETIC TRANSLATION of RNA into POLYPEPTIDES.
A DNA-dependent RNA polymerase present in bacterial, plant, and animal cells. The enzyme functions in the nucleolar structure and transcribes DNA into RNA. It has different requirements for cations and salts than RNA polymerase II and III and is not inhibited by alpha-amanitin. EC 2.7.7.6.
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 2.7.7.6.
Histochemical localization of immunoreactive substances using labeled antibodies as reagents.
The largest of polypeptide chains comprising immunoglobulins. They contain 450 to 600 amino acid residues per chain, and have molecular weights of 51-72 kDa.
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.
An enzyme that synthesizes DNA on an RNA template. It is encoded by the pol gene of retroviruses and by certain retrovirus-like elements. EC 2.7.7.49.
A DNA repair enzyme that catalyzes DNA synthesis during base excision DNA repair. EC 2.7.7.7.
DNA sequences which are recognized (directly or indirectly) and bound by a DNA-dependent RNA polymerase during the initiation of transcription. Highly conserved sequences within the promoter include the Pribnow box in bacteria and the TATA BOX in eukaryotes.
The sequential correspondence of nucleotides in one nucleic acid molecule with those of another nucleic acid molecule. Sequence homology is an indication of the genetic relatedness of different organisms and gene function.
Cells propagated in vitro in special media conducive to their growth. Cultured cells are used to study developmental, morphologic, metabolic, physiologic, and genetic processes, among others.
The parts of a transcript of a split GENE remaining after the INTRONS are removed. They are spliced together to become a MESSENGER RNA or other functional RNA.
Use of restriction endonucleases to analyze and generate a physical map of genomes, genes, or other segments of DNA.
DNA present in neoplastic tissue.
Established cell cultures that have the potential to propagate indefinitely.
The degree of similarity between sequences of amino acids. This information is useful for the analyzing genetic relatedness of proteins and species.
Biochemical identification of mutational changes in a nucleotide sequence.
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)
A technique that localizes specific nucleic acid sequences within intact chromosomes, eukaryotic cells, or bacterial cells through the use of specific nucleic acid-labeled probes.
Any of the processes by which nuclear, cytoplasmic, or intercellular factors influence the differential control (induction or repression) of gene action at the level of transcription or translation.
The relationships of groups of organisms as reflected by their genetic makeup.
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.
Domesticated bovine animals of the genus Bos, usually kept on a farm or ranch and used for the production of meat or dairy products or for heavy labor.
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.
The determination of the pattern of genes expressed at the level of GENETIC TRANSCRIPTION, under specific circumstances or in a specific cell.
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.
The proportion of one particular in the total of all ALLELES for one genetic locus in a breeding POPULATION.
Identification of proteins or peptides that have been electrophoretically separated by blot transferring from the electrophoresis gel to strips of nitrocellulose paper, followed by labeling with antibody probes.
Elements of limited time intervals, contributing to particular results or situations.
A mutation caused by the substitution of one nucleotide for another. This results in the DNA molecule having a change in a single base pair.
A group of deoxyribonucleotides (up to 12) in which the phosphate residues of each deoxyribonucleotide act as bridges in forming diester linkages between the deoxyribose moieties.
The ordered rearrangement of gene regions by DNA recombination such as that which occurs normally during development.
Studies determining the effectiveness or value of processes, personnel, and equipment, or the material on conducting such studies. For drugs and devices, CLINICAL TRIALS AS TOPIC; DRUG EVALUATION; and DRUG EVALUATION, PRECLINICAL are available.
The rate dynamics in chemical or physical systems.
Polypeptide chains, consisting of 211 to 217 amino acid residues and having a molecular weight of approximately 22 kDa. There are two major types of light chains, kappa and lambda. Two Ig light chains and two Ig heavy chains (IMMUNOGLOBULIN HEAVY CHAINS) make one immunoglobulin molecule.
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.
Laboratory techniques that involve the in-vitro synthesis of many copies of DNA or RNA from one original template.
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.
The infiltrating of tissue specimens with paraffin, as a supporting substance, to prepare for sectioning with a microtome.
Excrement from the INTESTINES, containing unabsorbed solids, waste products, secretions, and BACTERIA of the DIGESTIVE SYSTEM.
Detection of RNA that has been electrophoretically separated and immobilized by blotting on nitrocellulose or other type of paper or nylon membrane followed by hybridization with labeled NUCLEIC ACID PROBES.
Proteins prepared by recombinant DNA technology.
Endogenous substances, usually proteins, which are effective in the initiation, stimulation, or termination of the genetic transcription process.
The outward appearance of the individual. It is the product of interactions between genes, and between the GENOTYPE and the environment.
Infections produced by oncogenic viruses. The infections caused by DNA viruses are less numerous but more diverse than those caused by the RNA oncogenic viruses.
RNA present in neoplastic tissue.
A heat stable DNA-DIRECTED DNA POLYMERASE from the bacteria Thermus aquaticus. It is widely used for the amplification of genes through the process of POLYMERASE CHAIN REACTION. EC 2.7.7.-.
The process by which a DNA molecule is duplicated.
A type of chromosome aberration characterized by CHROMOSOME BREAKAGE and transfer of the broken-off portion to another location, often to a different chromosome.
An enzyme that catalyses RNA-template-directed extension of the 3'- end of an RNA strand by one nucleotide at a time, and can initiate a chain de novo. (Enzyme Nomenclature, 1992, p293)
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.
Diseases of the domestic dog (Canis familiaris). This term does not include diseases of wild dogs, WOLVES; FOXES; and other Canidae for which the heading CARNIVORA is used.
Genotypic differences observed among individuals in a population.
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.
Hybridization of a nucleic acid sample to a very large set of OLIGONUCLEOTIDE PROBES, which have been attached individually in columns and rows to a solid support, to determine a BASE SEQUENCE, or to detect variations in a gene sequence, GENE EXPRESSION, or for GENE MAPPING.
Proteins which bind to DNA. The family includes proteins which bind to both double- and single-stranded DNA and also includes specific DNA binding proteins in serum which can be used as markers for malignant diseases.
The functional hereditary units of BACTERIA.
Removal and pathologic examination of specimens in the form of small pieces of tissue from the living body.
Electrophoresis in which a polyacrylamide gel is used as the diffusion medium.
The parts of a macromolecule that directly participate in its specific combination with another molecule.
Cells grown in vitro from neoplastic tissue. If they can be established as a TUMOR CELL LINE, they can be propagated in cell culture indefinitely.
Proteins found in any species of bacterium.
Deoxyribonucleic acid that makes up the genetic material of fungi.
A positive regulatory effect on physiological processes at the molecular, cellular, or systemic level. At the molecular level, the major regulatory sites include membrane receptors, genes (GENE EXPRESSION REGULATION), mRNAs (RNA, MESSENGER), and proteins.
MOLECULAR BIOLOGY techniques used in the diagnosis of disease.
Polymers made up of a few (2-20) nucleotides. In molecular genetics, they refer to a short sequence synthesized to match a region where a mutation is known to occur, and then used as a probe (OLIGONUCLEOTIDE PROBES). (Dorland, 28th ed)
Extrachromosomal, usually CIRCULAR DNA molecules that are self-replicating and transferable from one organism to another. They are found in a variety of bacterial, archaeal, fungal, algal, and plant species. They are used in GENETIC ENGINEERING as CLONING VECTORS.
A latent susceptibility to disease at the genetic level, which may be activated under certain conditions.
A phenotypically recognizable genetic trait which can be used to identify a genetic locus, a linkage group, or a recombination event.
Enzymes that catalyze the transfer of multiple ADP-RIBOSE groups from nicotinamide-adenine dinucleotide (NAD) onto protein targets, thus building up a linear or branched homopolymer of repeating ADP-ribose units i.e., POLY ADENOSINE DIPHOSPHATE RIBOSE.
Proteins found in any species of virus.
The process in which substances, either endogenous or exogenous, bind to proteins, peptides, enzymes, protein precursors, or allied compounds. Specific protein-binding measures are often used as assays in diagnostic assessments.
Diseases of domestic cattle of the genus Bos. It includes diseases of cows, yaks, and zebus.
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.
Any of the processes by which nuclear, cytoplasmic, or intercellular factors influence the differential control of gene action in neoplastic tissue.
Ordered rearrangement of B-lymphocyte variable gene regions of the IMMUNOGLOBULIN HEAVY CHAINS, thereby contributing to antibody diversity. It occurs during the first stage of differentiation of the IMMATURE B-LYMPHOCYTES.
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).
Deoxyribonucleic acid that makes up the genetic material of helminths.
Virus diseases caused by the HERPESVIRIDAE.
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.
The uptake of naked or purified DNA by CELLS, usually meaning the process as it occurs in eukaryotic cells. It is analogous to bacterial transformation (TRANSFORMATION, BACTERIAL) and both are routinely employed in GENE TRANSFER TECHNIQUES.
A large collection of DNA fragments cloned (CLONING, MOLECULAR) from a given organism, tissue, organ, or cell type. It may contain complete genomic sequences (GENOMIC LIBRARY) or complementary DNA sequences, the latter being formed from messenger RNA and lacking intron sequences.
Remnant of a tumor or cancer after primary, potentially curative therapy. (Dr. Daniel Masys, written communication)
A genetic rearrangement through loss of segments of DNA or RNA, bringing sequences which are normally separated into close proximity. This deletion may be detected using cytogenetic techniques and can also be inferred from the phenotype, indicating a deletion at one specific locus.
A cell line derived from cultured tumor cells.
The record of descent or ancestry, particularly of a particular condition or trait, indicating individual family members, their relationships, and their status with respect to the trait or condition.
Compounds and molecular complexes that consist of very large numbers of atoms and are generally over 500 kDa in size. In biological systems macromolecular substances usually can be visualized using ELECTRON MICROSCOPY and are distinguished from ORGANELLES by the lack of a membrane structure.
A country spanning from central Asia to the Pacific Ocean.
The status during which female mammals carry their developing young (EMBRYOS or FETUSES) in utero before birth, beginning from FERTILIZATION to BIRTH.
Constituent of 30S subunit prokaryotic ribosomes containing 1600 nucleotides and 21 proteins. 16S rRNA is involved in initiation of polypeptide synthesis.
A specific pair of GROUP D CHROMOSOMES of the human chromosome classification.
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.
DNA sequences encoding RIBOSOMAL RNA and the segments of DNA separating the individual ribosomal RNA genes, referred to as RIBOSOMAL SPACER DNA.
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).
An individual having different alleles at one or more loci regarding a specific character.
A large lobed glandular organ in the abdomen of vertebrates that is responsible for detoxification, metabolism, synthesis and storage of various substances.
The total number of cases of a given disease in a specified population at a designated time. It is differentiated from INCIDENCE, which refers to the number of new cases in the population at a given time.
The larger subunits of MYOSINS. The heavy chains have a molecular weight of about 230 kDa and each heavy chain is usually associated with a dissimilar pair of MYOSIN LIGHT CHAINS. The heavy chains possess actin-binding and ATPase activity.
A category of nucleic acid sequences that function as units of heredity and which code for the basic instructions for the development, reproduction, and maintenance of organisms.
Infections with unicellular organisms formerly members of the subkingdom Protozoa. The infections may be experimental or veterinary.
Sequences of DNA in the genes that are located between the EXONS. They are transcribed along with the exons but are removed from the primary gene transcript by RNA SPLICING to leave mature RNA. Some introns code for separate genes.
Individuals whose ancestral origins are in the southeastern and eastern areas of the Asian continent.
Diseases of domestic swine and of the wild boar of the genus Sus.
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).
Immunologic techniques based on the use of: (1) enzyme-antibody conjugates; (2) enzyme-antigen conjugates; (3) antienzyme antibody followed by its homologous enzyme; or (4) enzyme-antienzyme complexes. These are used histologically for visualizing or labeling tissue specimens.
Any method used for determining the location of and relative distances between genes on a chromosome.
A specific pair of GROUP E CHROMOSOMES of the human chromosome classification.
Models used experimentally or theoretically to study molecular shape, electronic properties, or interactions; includes analogous molecules, computer-generated graphics, and mechanical structures.
Diseases of domestic and wild horses of the species Equus caballus.
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.
Ordered rearrangement of T-cell variable gene regions coding for the gamma-chain of antigen receptors.
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.
The sum of the weight of all the atoms in a molecule.
The domestic dog, Canis familiaris, comprising about 400 breeds, of the carnivore family CANIDAE. They are worldwide in distribution and live in association with people. (Walker's Mammals of the World, 5th ed, p1065)
Transport proteins that carry specific substances in the blood or across cell membranes.
Sudden increase in the incidence of a disease. The concept includes EPIDEMICS and PANDEMICS.
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.
The technique of using FIXATIVES in the preparation of cytologic, histologic, or pathologic specimens for the purpose of maintaining the existing form and structure of all the constituent elements.
Virus infections caused by the PARVOVIRIDAE.
Mature LYMPHOCYTES and MONOCYTES transported by the blood to the body's extravascular space. They are morphologically distinguishable from mature granulocytic leukocytes by their large, non-lobed nuclei and lack of coarse, heavily stained cytoplasmic granules.
Neoplasms of the skin and mucous membranes caused by papillomaviruses. They are usually benign but some have a high risk for malignant progression.
The type species of LYMPHOCRYPTOVIRUS, subfamily GAMMAHERPESVIRINAE, infecting B-cells in humans. It is thought to be the causative agent of INFECTIOUS MONONUCLEOSIS and is strongly associated with oral hairy leukoplakia (LEUKOPLAKIA, HAIRY;), BURKITT LYMPHOMA; and other malignancies.
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).
A form of fluorescent antibody technique utilizing a fluorochrome conjugated to an antibody, which is added directly to a tissue or cell suspension for the detection of a specific antigen. (Bennington, Saunders Dictionary & Encyclopedia of Laboratory Medicine and Technology, 1984)
A strain of albino rat used widely for experimental purposes because of its calmness and ease of handling. It was developed by the Sprague-Dawley Animal Company.
An infant during the first month after birth.
Infection with CYTOMEGALOVIRUS, characterized by enlarged cells bearing intranuclear inclusions. Infection may be in almost any organ, but the salivary glands are the most common site in children, as are the lungs in adults.
A single nucleotide variation in a genetic sequence that occurs at appreciable frequency in the population.
Infections with bacteria of the genus CHLAMYDIA.
Deletion of sequences of nucleic acids from the genetic material of an individual.
Naturally occurring or experimentally induced animal diseases with pathological processes sufficiently similar to those of human diseases. They are used as study models for human diseases.
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)
Proteins which are found in membranes including cellular and intracellular membranes. They consist of two types, peripheral and integral proteins. They include most membrane-associated enzymes, antigenic proteins, transport proteins, and drug, hormone, and lectin receptors.
An individual in which both alleles at a given locus are identical.
The first continuously cultured human malignant CELL LINE, derived from the cervical carcinoma of Henrietta Lacks. These cells are used for VIRUS CULTIVATION and antitumor drug screening assays.
Procedures for collecting, preserving, and transporting of specimens sufficiently stable to provide accurate and precise results suitable for clinical interpretation.
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.
A mammalian fetus expelled by INDUCED ABORTION or SPONTANEOUS ABORTION.
Constituent of the 40S subunit of eukaryotic ribosomes. 18S rRNA is involved in the initiation of polypeptide synthesis in eukaryotes.
A family of enveloped, linear, double-stranded DNA viruses infecting a wide variety of animals. Subfamilies, based on biological characteristics, include: ALPHAHERPESVIRINAE; BETAHERPESVIRINAE; and GAMMAHERPESVIRINAE.
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.
Any of the processes by which nuclear, cytoplasmic, or intercellular factors influence the differential control of gene action in enzyme synthesis.
A type of IN SITU HYBRIDIZATION in which target sequences are stained with fluorescent dye so their location and size can be determined using fluorescence microscopy. This staining is sufficiently distinct that the hybridization signal can be seen both in metaphase spreads and in interphase nuclei.
A prediction of the probable outcome of a disease based on a individual's condition and the usual course of the disease as seen in similar situations.
A species of the genus SACCHAROMYCES, family Saccharomycetaceae, order Saccharomycetales, known as "baker's" or "brewer's" yeast. The dried form is used as a dietary supplement.
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)
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.
A negative regulatory effect on physiological processes at the molecular, cellular, or systemic level. At the molecular level, the major regulatory sites include membrane receptors, genes (GENE EXPRESSION REGULATION), mRNAs (RNA, MESSENGER), and proteins.
Observation of a population for a sufficient number of persons over a sufficient number of years to generate incidence or mortality rates subsequent to the selection of the study group.
The complete genetic complement contained in a DNA or RNA molecule in a virus.
Virulent bacteriophage and type species of the genus T7-like phages, in the family PODOVIRIDAE, that infects E. coli. It consists of linear double-stranded DNA, terminally redundant, and non-permuted.
Type species of CHLAMYDIA causing a variety of ocular and urogenital diseases.
Addition of methyl groups to DNA. DNA methyltransferases (DNA methylases) perform this reaction using S-ADENOSYLMETHIONINE as the methyl group donor.
Infections with species of the genus MYCOPLASMA.
Diseases of the domestic cat (Felis catus or F. domesticus). This term does not include diseases of the so-called big cats such as CHEETAHS; LIONS; tigers, cougars, panthers, leopards, and other Felidae for which the heading CARNIVORA is used.
Techniques used in studying bacteria.
3 beta,12 beta,14-Trihydroxy-5 beta-card-20(22)-enolide. A cardenolide which is the aglycon of digoxin. Can be obtained by hydrolysis of digoxin or from Digitalis orientalis L. and Digitalis lanata Ehrh.
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.
The property of objects that determines the direction of heat flow when they are placed in direct thermal contact. The temperature is the energy of microscopic motions (vibrational and translational) of the particles of atoms.
Death resulting from the presence of a disease in an individual, as shown by a single case report or a limited number of patients. This should be differentiated from DEATH, the physiological cessation of life and from MORTALITY, an epidemiological or statistical concept.
A variety of simple repeat sequences that are distributed throughout the GENOME. They are characterized by a short repeat unit of 2-8 basepairs that is repeated up to 100 times. They are also known as short tandem repeats (STRs).
An enzyme that catalyzes the synthesis of polyadenylic acid from ATP. May be due to the action of RNA polymerase (EC 2.7.7.6) or polynucleotide adenylyltransferase (EC 2.7.7.19). EC 2.7.7.19.

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 - Amplified Fragment Length Polymorphism;Breed Discrimination;Chicken;Polymerase Chain Reaction;Nucleotide Polymorphism;
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 ...
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.
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
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 ...
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 ...
[110] 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.
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. ...
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 ...
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 ...
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
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 fluid of women accessing a sexually transmitted disease clinic. Sexually Transmitted Diseases, 33, 22-25. doi10.1097/01.olq.0000194600.83825.81
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, ...
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, ...
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 ...
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.
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)
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.
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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.
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 IPFS - polymerase chain reaction the many methods now used to rapid and widespread application as the polymerase chain reaction
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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 ...
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 ...
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 ...
Read independent reviews on High Pure PCR Template Preparation Kit from Roche Applied Science - a member of the Roche Group on SelectScience
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 ...
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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 ...
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This realization is quite discouraging; I have followed doctors orders to a T, I have taken every dose of Sprycel, on time for the past year, and I am exactly where I started. I am at square one, taking more medication than I had hoped to be taking at this point. I now look back and wonder whether or not I should have questioned my very first doctor when he started my treatment with the highest, possible dose of Sprycel. He was not a CML specialist, so he was simply reading the guidelines for someone in the acute phase of chronic myelogenous leukemia, not the chronic phase, which it was determined, despite my 385,000 white cell count, that I was in. Maybe he knew what he was doing ...
Chinese officials claimed yesterday that COVID-19 tests on 10.9 million people in Qingdao City, Shandong Province, had all returned negative results, but Taiwans Health Minister, Chen Shih-chung (陳時中), said today that such a result was impossible. Citywide tests for Qingdao Citys population of 11 million were ordered after a cluster infection of 13 people centered around a local hospital was discovered recently. The cluster was claimed to be Chinas first case of locally transmitted COVID-19 in over two months. The. Read more ...
BACKGROUND: The interaction between host lymphocytes and endothelial cells on the transplanted organ is believed to play an important role in acute and chronic graft rejection. Trafficking and recruitment of lymphocytes to the site of inflammation is known to be controlled by several cytokines and chemokines. It is unclear whether endothelial cells themselves can be a source of inflammatory chemoattractant molecules on alloimmune induction. METHODS: Using a semiquantitative polymerase chain reaction method, the authors analyzed the expression of chemokine mRNA coding for interferon (IFN)-gamma-induced protein 10 (IP-10) and monokine induced by IFN-gamma (Mig) in a pool of human aortic endothelial cells. Both of these chemokines are known to be induced by IFN-gamma. Endothelial cell-derived chemokine mRNA was assayed at rest, after IFN-gamma activation, and after co-culture with allogeneic peripheral blood mononuclear cells (PBMC) from normal blood donors with and without a monoclonal antibody to IFN
Polymerase Chain Reaction Technique: Introduction, Mechanism, Steps, and Objectives Polymerase chain reaction( PCR) is a laboratory technique used to make multiple copies of a segment of DNA. PCR is extremely accurate and ca
With the increasing incidence and mortality of fungal infection, the requirements for strict diagnostic approaches became a very urgent issue. Because of the traditional detective techniques, such as culture, gave poor diagnostic outcomes, the molecular biological techniques are expected to develop the potential diagnostic approaches. During the past decades, we have carried out serial studies on the molecular properties of pathogenic fungi, and we would like to review as following. Firstly, we applied several molecular tools in classification and identification of pathogenic fungi. We performed random amplification of polymorphic DNA (RAPD), restriction fragment length polymorphism (RFLP) and other techniques in studying the typing, to classify and identify the properties of Dermatophytes, Candida spp., Crypotococcus neoformans, Dematiaceous fungi, and Aspergillus spp. Interestingly, we found the same T. rubrum strain might infect different sites of the host, while a site-specificity displayed in T.
The primer and amplicon length have been found to affect PCR based estimates of microbial diversity by pyrosequencing, while other PCR conditions have not been addressed using any deep sequencing method. The present study determined the effects of polymerase, template dilution and PCR cycle number using the Solexa platform. The PfuUltra II Fusion HS DNA Polymerase (Stratagene) with higher fidelity showed lower amount of PCR artifacts and determined lower taxa richness than the Ex Taq (Takara). More importantly, the two polymerases showed different efficiencies for amplifying some of very abundant sequences, and determined significantly different community structures. As expected, the dilution of the DNA template resulted in a reduced estimation of taxa richness, particularly at the 200 fold dilution level, but the community structures were similar for all dilution levels. The 30 cycle group increased the PCR artifacts while comparing to the 25 cycle group, but the determined taxa richness was lower than
Roche Diagnostics has launched the new LightCycler Nano real-time PCR system for use in research in the US. Next to its fancy design it is also one of the
PCR product size - posted in PCR, RT-PCR and Real-Time PCR: I have a forward primer started from nucleotide no. 79 till 99 and a reverse primer located at nucleotide no. 114 till 391. From there, how can I predict my RT-PCR product size (from cDNA)? I have designed my primer gDNA sequence where there is an intron within my primer design. Should I exclude or include the intron sequence in order to count my PCR producr size?
RECOMMENDED: If you have Windows errors then we strongly recommend that you download and run this (Windows) Repair Tool.. Sep 24, 2011. Error-prone polymerase chain reactions (epPCRs) are often used to introduce mutations in random mutagenesis, which has been used as a.. Random mutagenesis by error-prone PCR. nucleotide mutations into a parent sequence that takes advantage of the polymerase chain reaction (PCR).. Sql Error Logs Sql Server 2005 Mar 24, 2015. Here are various ways to find the SQL Server ErrorLog location. In SQL Server 2005, we would see a key name in the format of MSSQL.n. MySQL Database for Global Regions Starting from $10.88/mo. More Deals from Oct24 Over the weekend a website I run stopped functioning, recording the following error in. Nature Reviews Drug Discovery - In the mammalian immune system, antibodies undergo affinity maturation and. an antibody library with mutations within the variable genes either by error-prone polymerase chain reaction, E. coli mutator strains or ...
Work package 2 (WP2) is headed by Prof. Heribert Insam from the Institute of Microbiology at the University of Innsbruck in collaboration with the Department of Agrifood and Environmental Sciences (DiSPAA) of the University of Florence (IT). The general aim of WP2 is to gain an in-depth understanding of the structure and function of microbial communities involved in deadwood decomposition along the selected climosequences. Molecular methods based on polymerase chain reaction (PCR) amplification of rRNA genes allow the profiling of complex microbial communities on the basis of sequence diversity. At the selected sites, archaeal, fungal and bacterial diversity are being analysed by community level genetic fingerprinting (DGGE, denaturing gradient gel electrophoresis profiles) based on total community DNA. A further characterisation of soil microbiota is being assessed by using specific primer sets targeting functional genes involved in key processes of C and N cycles. PCR-DGGE analyses are being ...
A multiplex polymerase chain reaction protocol for the simultaneous analysis of the glutathione S-transferase GSTM1 and GSTT1 polymorphisms
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 ...
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
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 ...
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 ...
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
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 ...
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
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). 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. ...
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 ...
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
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 ...
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
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 ...
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).
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
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.. ...
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 ...
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... ...
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. ...
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: 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
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 ...
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 ...
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, ...
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 ...
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 by amplification refractory mutation ...
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 ...
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 ...
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
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 ...
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.. ...
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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 ...
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. ...
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 ...
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 - ...
Among newborns, CMV testing with DBS real-time PCR compared with saliva rapid culture had low sensitivity, limiting its value as a screening test.
PCR-Based Genotyping Methods. An introduction to PCR-RFLP/CAPS, and dCAPS. Common PCR-based Genotyping Methods for SNP Analysis. SNPs can have up to 4 alleles (A/C/G/T), but two alleles are most common . These methods can only positively detect one allele. PCR -RFLP / CAPS Slideshow 6726805 by tatiana-stanley
IL-10 inhibits the ability of macrophage but not B cell APC to stimulate cytokine synthesis by Th1 T cell clones. In this study we have examined the direct effects of IL-10 on both macrophage cell lines and normal peritoneal macrophages. LPS (or LPS and IFN-gamma)-induced production of IL-1, IL-6, and TNF-alpha proteins was significantly inhibited by IL-10 in two macrophage cell lines. Furthermore, IL-10 appears to be a more potent inhibitor of monokine synthesis than IL-4 when added at similar concentrations. LPS or LPS- and IFN-gamma-induced expression of IL-1 alpha, IL-6, or TNF-alpha mRNA was also inhibited by IL-10 as shown by semiquantitative polymerase chain reaction or Northern blot analysis. Inhibition of LPS-induced IL-6 secretion by IL-10 was less marked in FACS-purified peritoneal macrophages than in the macrophage cell lines. However, IL-6 production by peritoneal macrophages was enhanced by addition of anti-IL-10 antibodies, implying the presence in these cultures of endogenous ...
To generate reasonable serum concentrations of interferon the conventional interferons need to be administered at least three times a week and this frequent administration is uncomfortable and inconvenient for the patient. Even with thrice weekly injections of conventional interferon there is still marked fluctuation of the serum concentration of interferon and previous attempts to administer the drug more often to reduce these wide variations were only partially successful. To resolve the problems associated with the short half-life of conventional interferons the interferon protein has been attached to a second molecule, polyethylene glycol, to produce much larger, more stable compounds. Immunohistochemical staining has no role to play in the routine assessment of liver biopsies in patients with hepatitis C because no commercially available antibody has been shown to work reliably on formalin-fixed, paraffin-embedded tissue. In situ polymerase chain reaction (PCR) to detect the viral RNA in ...
A polymerase chain reaction (PCR)-based diagnostic assay was developed that rapidly and reliably differentiates the sibling species of the Anopheles claviger complex, An. claviger s.s. and An. petragnani. The assay makes use of nucleotide differences in the internal transcribed spacer 2 ribosomal DNA sequences to generate PCR products of specific length for each of the two species. In evaluating the test, 580 of 592 field-collected An. claviger s.l. specimens were unambiguously identified as one of the two sibling species. Due to poor DNA quality, the remaining 12 specimens yielded no PCR product. Of the 592 mosquitoes, 407 larval specimens had been identified morphologically prior to species-specific DNA amplification, and in all instances PCR identification corroborated with morphologic identification. Mosquitoes identified as An. claviger s.s. came from various localities all over Europe and from Israel. Those identified as An. petragnani were collected in southern France and Spain. The species
The clustered regularly interspaced short palindromic repeats-CRISPR-associated protein 9 (CRISPR-Cas9) system is considered as a technological revolution in targeted mutagenesis. However, a large amount of time and cost is needed to screen for the CRISPR-Cas9 induced mutants from a usual large number of initial samples. Thus, Chun Wang and Kejian Wang from the Chinese Academy of Agricultural Sciences presented a cost-effective and sensitive screening technique for identifying mutants based on conventional polymerase chain reaction (PCR). They called this new technique as annealing at critical temperature PCR (ACT-PCR). ACT-PCR needs only one PCR step and then execution of agarose gel electrophoresis. Because of its simplicity, ACT-PCR is suitable for rapid, large-scale screening of CRISPR-Cas9-induced mutants. Read more description about ACT-PCR in Plant Genome Editing with CRISPR Systems.. ...
Young children are more likely to become infected by respiratory viruses and experience severe symptoms and longer duration of illness than other age-groups, according to a study yesterday in Clinical Infectious Diseases.. The University of Utahs Utah Better Identification of Germs-Longitudinal Viral Epidemiology (BIG-LoVE) study analyzed symptom reports and nasal swabs from 26 households comprising 108 individuals over 52 weeks (4,166 person-weeks). Participants ranged in age from 1 day to 57 years, and 85% of households had at least one child.. Researchers used polymerase chain reaction (PCR) testing to detect 16 respiratory viruses from 2009 to 2010, a period that included the second wave of the 2009 H1N1 pandemic.. Respiratory viruses were detected 783 times in participants over the year, and 440 (56%) of those infections were symptomatic. Individuals had an average of five respiratory viruses per year.. Children under age 5 tested positive for respiratory viruses in 50% of person-weeks, ...
Phytopathology 92:112-116...Phytopathology 92:112-116...Simultaneous One-Tube Quantification of Host and Pathogen DNA with Real-Time Polymerase Chain Reaction...L. M. Winton , J. K. Stone , L. S. Watrud , and E. M. Hansen...
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: ...
"Polymerase chain reaction makes billions of DNA copies". WhatisBiotechnology.org. Retrieved 2020-12-05. v t e. ... During his postdoctoral career in the United States, he put forward the idea of the polymerase chain reaction (PCR). Kleppe ... Repair replications of short synthetic DNA's as catalyzed by DNA polymerases". Journal of Molecular Biology. 56 (2): 341-61. ... "process called repair replication for synthesizing short DNA duplexes and single-stranded DNA by polymerases." Because of his ...
by using polymerase chain reaction and gene probes". J. Clin. Microbiol. 30 (1): 74-8. doi:10.1128/JCM.30.1.74-78.1992. PMC ... Polymerase Chain Reactions) have been developed to detect specific Giardia spp. Gene probe-based detection is also used to ...
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 ... The device has a thermal block with holes where tubes holding the reaction mixtures can be inserted. The cycler then raises and ... This led to a cumbersome machine based on an automated pipettor, with open reaction tubes. Later, the PCR process was adapted ...
Amplification is done using polymerase chain reactions or PCR. The choice of primers can allow either for a single gene to be ... These secondary structures cause somatic mosaicism by lagging DNA polymerase in Okazaki fragments and by disrupting DNA ... in which DNA is used to generate a complementary RNA strand by RNA polymerase, and translation, in which RNA is used to produce ...
Abdul Khaliq, R; Kafafy, R.; Salleh, H. M.; Faris, W. F. (2012). "Enhancing the efficiency of polymerase chain reaction using ... are linked to the nanostructure via short peptide chains. Injected intravenously, the graphene strips with the drug payload ... "N-Doping of Graphene Through Electrothermal Reactions with Ammonia". Science. 324 (5928): 768-71. Bibcode:2009Sci...324..768W. ... at the University of Western Australia discovered nano sized fragments of graphene can speed up the rate of chemical reactions ...
Another method of screening is with polymerase chain reaction (PCR). Some libraries are stored as pools of clones and screening ...
Polymerase chain reaction (PCR) has been used on tissue specimens. Cryptococcosis can rarely occur in the non-immunosuppressed ... a higher mortality rate in patients with non-HIV cryptococcal meningitis secondary to the role of T-cell mediated reaction and ...
"Isolation of telomere junction fragments by anchored polymerase chain reaction". Proceedings. Biological Sciences. 247 (1318): ...
Nirenberg and Matthaei experiment Polymerase chain reaction optimization Gregorio, Nicole E.; Levine, Max Z.; Oza, Javin P. ( ... The openness of the reaction is ideal for inserting the modified tRNAs and unnatural amino acids required for such a reaction. ... In contrast, once DNA is inserted into live cells, the reaction cannot be accessed until it is over and the cells are lysed. ... Common components of a cell-free reaction include a cell extract, an energy source, a supply of amino acids, cofactors such as ...
Polymerase chain reaction[edit]. 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[edit]. 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[edit]. 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 ...
Other methods include culture and polymerase chain reaction, but these are not routinely available and the results do not ... Tay ST, Nazma S, Rohani MY (1996). "Diagnosis of scrub typhus in Malaysian aborigines using nested polymerase chain reaction". ... December 2006). "Rapid diagnosis of scrub typhus in rural Thailand using polymerase chain reaction". Am. J. Trop. Med. Hyg. 75 ... by nested polymerase chain reaction". J Med Entomol. 38 (2): 308-311. doi:10.1603/0022-2585-38.2.308. PMID 11296840. Seong SY, ...
Graphene based nanofluid has been found to enhance Polymerase chain reaction efficiency. Nanofluids in solar collectors is ... "Enhancing the efficiency of polymerase chain reaction using graphene nanoflakes - Abstract - Nanotechnology - IOPscience". iop. ... The MWCNTs are functionalized in one pot using a free radical grafting reaction. The clove-functionalized MWCNTs are then ...
Polymerase chain reaction was used to amplify the amount of usable DNA. The United States Department of Defense had previously ...
Polymerase Chain Reaction was developed in 1980's by the scientist Kary Mullis. Mullis would later receive the Nobel Prize for ... "Principles and applications of polymerase chain reaction in medical diagnostic fields: a review". Brazilian Journal of ... A good example of this would be how Taq Polymerase was isolated from the bacteria Thermus aquaticus and was then used to make ... The small sample of the target DNA is added to a test tube along with DNA primers, DNA nucleotides, Taq Polymerase, and a ...
A study using polymerase chain reaction testing of stool samples suggested that symptomatic infection can exist even when ... of individuals infected when compared to polymerase chain reaction (PCR) testing. Reasons given for the failure of Direct ... "Detection of Blastocystis hominis in unpreserved stool specimens by using polymerase chain reaction". J. Parasitol. 92 (5): ... A study of IBS patients in the Middle East showed a "significantly increased" immune reaction in IBS patients to Blastocystis, ...
Polymerase chain reaction is a tool to amplify and detect specific DNA sequences. It can be used to help characterize the ...
Yokomi, Raymond K.; Mello, Alexandre F. S.; Saponari, Maria; Fletcher, Jacqueline (February 2008). "Polymerase Chain Reaction- ...
"Differential display of eukaryotic messenger RNA by means of the polymerase chain reaction". Science. 257 (5072): 967-71. PMID ...
December 2006). "Pneumolysin polymerase chain reaction for diagnosis of pneumococcal pneumonia and empyema in children". ... August 2006). "Diagnosis of Streptococcus pneumoniae meningitis by polymerase chain reaction amplification of the gene for ...
Method of diagnosing gummy stem blight in plants using a polymerase chain reaction assay. US 20010758073 Somai, B. M., Keinath ... Molecular tools such as Polymerase Chain Reaction (PCR), PCR-enzyme-linked immunosorbent assay and magnetic-capture ...
Polymerase chain reaction (DNA). DNA profiling is today possible with even very small quantities of blood: this is commonly ...
Polymerase chain reaction testing of the CSF does not show the presence of the parasite.[citation needed] The cause is ... The Mazzotti reaction, first described in 1948, is a symptom complex seen in patients after undergoing treatment of ... onchocerciasis with the medication diethylcarbamazine (DEC). Mazzotti reactions can be life-threatening, and are characterized ...
"Rapid identification of Fonsecaea by duplex polymerase chain reaction in isolates from patients with chromoblastomycosis". ...
January 2004). "Polymerase chain reaction-based genotype classification among human Blastocystis hominis populations isolated ...
Several different polymerase chain reaction (PCR) tests are available for the detection of Leishmania DNA. With this assay, a ... RNA polymerase II transcribes long polycistronic messages in the absence of defined RNA pol II promoters, and Leishmania has ...
One widely adopted WGA techniques is called degenerate oligonucleotide-primed polymerase chain reaction (DOP-PCR). This method ... usually Φ29 DNA polymerase, to accomplish the amplification of larger fragments and greater genome coverage than DOP-PCR. ...
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. ...
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)[6][23] 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.[236][237] ...
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,[24] which is similar ... The two RNA polymerases may recognize and bind to different kinds of promoters within the chloroplast genome.[35] The ribosomes ...
Many modern molecular tests such as flow cytometry, polymerase chain reaction (PCR), immunohistochemistry, cytogenetics, gene ...
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.[4]. A safe and effective vaccine against yellow fever exists, ...
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.[129] ... "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, ...
... 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 ...
In: Polymerase Chain Reaction (PCR) for Human Viral Diagnosis Clewley JP, ed. Boca Raton: CRC. pp 125-145. ...
α-1,4 glycogen chain)n + Pi ⇌ (α-1,4 glycogen chain)n-1 + α-D-glucose-1-phosphate.[2] ... 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 ...
"Phylogenetic analysis of Aquaspirillum magnetotacticum using polymerase chain reaction-amplified 16S rRNA-specific DNA". ...
... 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 - ...
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, ...
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).[30]. 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.[98] 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 ...
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.[2] ... "Classification and Nomenclature of Enzymes by the Reactions they Catalyse". 》International Union of Biochemistry and Molecular ...
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. • ...
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.[2] ... 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 ...
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 ...
Histone proteins are made up of long chains of amino acids. If the amino acids that are in the chain are changed, the shape of ... ribose polymerase) and its product poly(ADP)-ribose (PAR) accumulate at sites of DNA damage as part of a repair process.[32] ... The acetylation event converts the positively charged amine group on the side chain into a neutral amide linkage. This removes ... in which polymerase sensitivity allows for measuring methylation and other modifications as a DNA molecule is being sequenced.[ ...
Mullis, K.B; Faloona, F.L. (1987). "[21] Specific synthesis of DNA in vitro via a polymerase-catalyzed chain reaction". Methods ...
Next, a nearby tyrosine residue deprotonates the ε-amino group of the lysine residue.[10] The lysine chain then makes a ... In order for the reaction to proceed, S-Adenosyl methionine (SAM) and the lysine residue of the substrate histone tail must ... RNA polymerase control by chromatin structure. *Histone methylation. ReferencesEdit. *^ a b c d Wood A (2004). " ... transferring the methyl group to the lysine side chain. ...
The amino acids in a polypeptide chain are linked by peptide bonds. Once linked in the protein chain, an individual amino acid ... About 4,000 reactions are known to be catalysed by enzymes.[32] The rate acceleration conferred by enzymatic catalysis is often ... by proteins such as RNA polymerase. Most organisms then process the pre-mRNA (also known as a primary transcript) using various ... A linear chain of amino acid residues is called a polypeptide. A protein contains at least one long polypeptide. Short ...
Nested polymerase chain reaction involves two sets of primers, used in two successive runs of polymerase chain reaction, the ... Nested polymerase chain reaction (Nested PCR) is a modification of polymerase chain reaction intended to reduce non-specific ... Polymerase chain reaction itself is the process used to amplify DNA samples, via a temperature-mediated DNA polymerase. The ... The target DNA undergoes the first run of polymerase chain reaction with the first set of primers, shown in green. The ...
... 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 ...
Polymerase Chain Reaction (PCR) is one of the most important molecular diagnostic tools which allow the detection of nucleic ...
2006) Polymerase chain reaction detection of Y-chromosome sequences in vaginal fluid of women accessing a sexually transmitted ... 2006) Polymerase chain reaction detection of Y-chromosome sequences in vaginal fluid of women accessing a sexually transmitted ...
A novel temperature control method for shortening thermal cycling time to achieve rapid polymerase chain reaction (PCR) in a ... A novel temperature control method for shortening thermal cycling time to achieve rapid polymerase chain reaction (PCR) in a ... A novel temperature control method for shortening thermal cycling time to achieve rapid polymerase chain reaction (PCR) in a ... A novel temperature control method for shortening thermal cycling time to achieve rapid polymerase chain reaction (PCR) in a ...
The polymerase chain reaction method is used to quantify nucleic acids by amplifying a nucleic acid molecule with the enzyme ... Digital polymerase chain reaction (digital PCR, DigitalPCR, dPCR, or dePCR) is a biotechnological refinement of conventional ... "Polymerase Chain Reaction (PCR)". National Center for Biotechnology Information, U.S. National Library of Medicine. Duewer, ... Lee, S.Y.; Hwang, S.Y. (2015). "Application of digital polymerase chain reaction technology for noninvasive prenatal test". ...
... ( 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 ...
... 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 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 ...
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. ...
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., ...
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, 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 ...
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.. ...
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 ...
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 ...
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 ...
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 ...
... (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 ...
... (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 ...
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 ...
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 Burkholderia pseudomallei in blood samples using polymerase chain reaction.. Rattanathongkom A1, Sermswan RW, ... were used for amplification of bacterial DNA by the polymerase chain reaction (PCR). Amplification with these primers yielded a ...
... 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 ...
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. ...
... 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 ...
Recently, simple and rapid assays for quantifying mRNA expression by real-time reverse transcription-polymerase chain reaction ... CERVINE (CERVUS ELAPHUS) CYTOKINE mRNA QUANTIFICATION BY REAL-TIME POLYMERASE CHAIN REACTION. ... CYTOKINE mRNA QUANTIFICATION BY REAL-TIME POLYMERASE CHAIN REACTION," Journal of Wildlife Diseases 42(2), 219-233, (1 April ... CYTOKINE mRNA QUANTIFICATION BY REAL-TIME POLYMERASE CHAIN REACTION," Journal of Wildlife Diseases, 42(2), 219-233, (1 April ...
... 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 ...
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 ...
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. ...
A novel flow-through polymerase chain reaction (PCR) microfluidic system using vapor pressure was developed that can achieve ... A novel flow-through polymerase chain reaction (PCR) microfluidic system using vapor pressure was developed that can achieve ...
Polymerase Chain Reaction) is a cornerstone of molecular biology research. Constantly evolving PCR reagents and applications ... A mainstay of virtually every molecular biology lab, polymerase chain reaction (PCR) is an easy and affordable method for ...
DNA affinity columns that permits direct extraction of bacterial plasmids from a variety of foods for polymerase chain reaction ... DNA affinity columns that permits direct extraction of bacterial plasmids from a variety of foods for polymerase chain reaction ... Polymerase inhibitors present in both foods and enrichment media were removed efficiently.. ...
  • Nested polymerase chain reaction (Nested PCR) is a modification of polymerase chain reaction intended to reduce non-specific binding in products due to the amplification of unexpected primer binding sites. (wikipedia.org)
  • Flow immunophenotyping, DNA content analysis, and polymerase chain reaction (PCR) amplification for t(11;14) and t(14;18) were performed on 11 cases of typical mantle cell lymphoma (MCL), 5 cases of apparent MCL with proliferation centers (MCL-PC), and 5 cases of small lymphocytic lymphoma (SLL). (ashpublications.org)
  • 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). (elsevier.com)
  • Nested polymerase chain reaction involves two sets of primers, used in two successive runs of polymerase chain reaction, the second set intended to amplify a secondary target within the first run product. (wikipedia.org)
  • The target DNA undergoes the first run of polymerase chain reaction with the first set of primers, shown in green. (wikipedia.org)
  • The product from the first reaction undergoes a second run with the second set of primers, shown in red. (wikipedia.org)
  • Then, to perform PCR, the DNA template that contains the target is added to a tube that contains primers, free nucleotides, and an enzyme called DNA polymerase, and the mixture is placed in a PCR machine. (latestgkgs.com)
  • The DNA polymerase begins to synthesize new strands of DNA starting from the primers. (latestgkgs.com)
  • Low-level malaria infections detected by a sensitive polymerase chain reaction assay and use of this technique in the evaluation of malaria vaccines in an endemic area. (ox.ac.uk)
  • Polymerase Chain Reaction (PCR) is one of the most important molecular diagnostic tools which allow the detection of nucleic acid targets. (scialert.net)
  • Nucleic acid hybridisation and polymerase chain reaction in the the use of nucleic acid hybridisation and polymerase methods and does not, Application of polymerase chain reaction for detection of goats' milk adulteration by milk of cow - Volume 68 Issue 2 - JACEK BANIA, MACIEJ UGORSKI, ANTONI POLANOWSKI. (hochregal.net)
  • US7393644B2 Method for real-time detection of polymerase. (hochregal.net)
  • Detection of human papillomavirus type 16 DNA sequences in archival cervical tissues by the polymerase chain reaction. (ox.ac.uk)
  • We have evaluated the polymerase chain reaction for the detection of viral DNA sequences in paraffin-embedded archival tissues. (ox.ac.uk)
  • 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. (ox.ac.uk)
  • 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. (ox.ac.uk)
  • Global Polymerase Chain Reaction Technologies Industry valued approximately USD 7 million in 2016 is anticipated to grow with a healthy growth rate of more than 7% over the forecast period 2019-2025. (globalreportsstore.com)
  • Using barcodes and barcode databases, species specific Quantitative Polymerase Chain Reaction (qPCR) assays can be designed to detect the presence of target species where traditional sampling is inefficient or impossible. (confex.com)
  • The polymerase chain reaction Application-specific PCR F.A. Specific synthesis of DNA in vitro via a polymerase-catalyzed chain reaction. (hochregal.net)
  • The methods and systems may The polymerase chain reaction Application-specific PCR F.A. Specific synthesis of DNA in vitro via a polymerase-catalyzed chain reaction. (hochregal.net)
  • Polymerase chain reaction itself is the process used to amplify DNA samples, via a temperature-mediated DNA polymerase. (wikipedia.org)
  • Inverse polymerase chain reaction (Inverse PCR) is a variant of the polymerase chain reaction that is used to amplify DNA with only one known sequence. (wikipedia.org)
  • Then, like other polymerase chain reaction processes, the DNA is amplified by the temperature-sensitive DNA polymerase: A target region with an internal section of known sequence and unknown flanking regions is identified Genomic DNA is digested into fragments of a few kilobases by a usually low-moderate frequency (6-8 base) cutting restriction enzyme. (wikipedia.org)
  • Digital Polymerase Chain Reaction (dPCR) is an advancement of traditional polymerase chain reaction (PCR). (bigmarketresearch.com)
  • 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 disease, and increasing number of diagnostic centers across the globe. (bigmarketresearch.com)
  • Polymerase chain reaction( PCR) is a laboratory technique used to make multiple copies of a segment of DNA. (latestgkgs.com)
  • A novel temperature control method for shortening thermal cycling time to achieve rapid polymerase chain reaction (PCR) in a disposable polymer microfluidic device. (dtu.dk)
  • 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. (dtu.dk)
  • Polymerase Chain Reaction size and sequence by enzymatic method and cycling condition. (hochregal.net)
  • Learn about PCR (polymerase chain reaction) a method of analyzing a short sequence of DNA or RNA. (hochregal.net)
  • Polymerase Chain Reaction Validation, Optimization, Limitations and Applications The method is useful for coding the genetic sequence of various disease, Polymerase Chain Reaction (PCR) In Medical Application: An Analytical Report 2014-2020 - The global PCR market is projected to reach around US$27.4 billion by 2015. (hochregal.net)
  • 1/03/2009В В· Principles and applications of polymerase chain reaction in a broad range of medical diagnostic fields, methods, Polymerase Chain Reaction, You have free access to this content The polymerase chain reaction and its application in occupational and environmental medicine [Biomonitoring Methods, 2004]. (hochregal.net)
  • Un examen rétrospectif des dossiers pré- et post-intervention a servi à décrire le séjour à l'hôpital des patients ayant reçu un diagnostic de BAC-SA avant et après la mise en place du système de diagnostic de la PCR. (cjhp-online.ca)
  • Au total, 98 et 99 patients ont satisfait au critère d'inclusion respectivement avant et après la mise en place du système de diagnostic de la PCR. (cjhp-online.ca)
  • Polymerase Chain Reaction invented a method of amplifying any DNA region through repeated cycles of implications for research applications and in forensics. (hochregal.net)
  • The amount of product from the PCR increases with the number of temperature cycles that the reaction is subjected to. (wikipedia.org)
  • 17/04/2014В В· Polymerase Chain Reaction (PCR) Steps, Requirements and Applications biologyexams4u. (hochregal.net)
  • Methods Enzymol https://simple.wikipedia.org/wiki/Polymerase_chain_reaction Polymerase Chain Reaction. (hochregal.net)
  • PCR Methods Appl 1, 17 Genetic applications of an inverse polymerase chain reaction. (hochregal.net)
  • Methods and systems for polymerase chain reactions (PCR) that are capable of detecting amplified DNA during or after the PCR process. (hochregal.net)
  • 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, and data from 20 individual singleplex quantitative PCR reactions. (elsevier.com)
  • This powerful method has numerous applications in Theory and applications of the polymerase chain Theory and applications of the polymerase chain reaction. (hochregal.net)
  • 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. (ox.ac.uk)
  • Emphasis is placed on non-invasive analysis of single cell metabolites and the direct analysis of RNA and DNA from single cells, with a focus on polymerase chain reaction and fluorescence in situ hybridization. (moluna.de)
  • In MCL-PC, the mean intensity of IgM, Ig light chain, and CD20 expression was intermediate to the intensities observed in MCL and SLL. (ashpublications.org)
  • PDF) Polymerase chain reaction Theory practice and. (hochregal.net)
  • 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. (dsportstimes.com)
  • 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. (dsportstimes.com)
  • For a global outreach, the Polymerase Chain Reaction (PCR) Products study also classifies the market into a global distribution where key market demographics are established based on the majority of the market share. (dsportstimes.com)