Polymerase Chain Reaction: In vitro method for producing large amounts of specific DNA or RNA fragments of defined length and sequence from small amounts of short oligonucleotide flanking sequences (primers). The essential steps include thermal denaturation of the double-stranded target molecules, annealing of the primers to their complementary sequences, and extension of the annealed primers by enzymatic synthesis with DNA polymerase. The reaction is efficient, specific, and extremely sensitive. Uses for the reaction include disease diagnosis, detection of difficult-to-isolate pathogens, mutation analysis, genetic testing, DNA sequencing, and analyzing evolutionary relationships.Reverse Transcriptase Polymerase Chain Reaction: 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.Base Sequence: The sequence of PURINES and PYRIMIDINES in nucleic acids and polynucleotides. It is also called nucleotide sequence.Molecular Sequence Data: Descriptions of specific amino acid, carbohydrate, or nucleotide sequences which have appeared in the published literature and/or are deposited in and maintained by databanks such as GENBANK, European Molecular Biology Laboratory (EMBL), National Biomedical Research Foundation (NBRF), or other sequence repositories.DNA-Directed DNA Polymerase: DNA-dependent DNA polymerases found in bacteria, animal and plant cells. During the replication process, these enzymes catalyze the addition of deoxyribonucleotide residues to the end of a DNA strand in the presence of DNA as template-primer. They also possess exonuclease activity and therefore function in DNA repair.RNA, Messenger: 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.DNA Primers: 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.Real-Time Polymerase Chain Reaction: Methods used for detecting the amplified DNA products from the polymerase chain reaction as they accumulate instead of at the end of the reaction.RNA Polymerase II: 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.Sensitivity and Specificity: 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)DNA, Viral: Deoxyribonucleic acid that makes up the genetic material of viruses.Amino Acid Sequence: 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.DNA, Bacterial: Deoxyribonucleic acid that makes up the genetic material of bacteria.DNA: 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).Cloning, Molecular: 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.DNA-Directed RNA Polymerases: Enzymes that catalyze DNA template-directed extension of the 3'-end of an RNA strand one nucleotide at a time. They can initiate a chain de novo. In eukaryotes, three forms of the enzyme have been distinguished on the basis of sensitivity to alpha-amanitin, and the type of RNA synthesized. (From Enzyme Nomenclature, 1992).Transcription, Genetic: The biosynthesis of RNA carried out on a template of DNA. The biosynthesis of DNA from an RNA template is called REVERSE TRANSCRIPTION.Genotype: The genetic constitution of the individual, comprising the ALLELES present at each GENETIC LOCUS.DNA Polymerase I: A DNA-dependent DNA polymerase characterized in prokaryotes and may be present in higher organisms. It has both 3'-5' and 5'-3' exonuclease activity, but cannot use native double-stranded DNA as template-primer. It is not inhibited by sulfhydryl reagents and is active in both DNA synthesis and repair. EC 2.7.7.7.Blotting, Southern: 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.Oligonucleotide 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.Mutation: 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.Gene Expression: The phenotypic manifestation of a gene or genes by the processes of GENETIC TRANSCRIPTION and GENETIC TRANSLATION.DNA Polymerase II: A DNA-dependent DNA polymerase characterized in E. coli and other lower organisms. It may be present in higher organisms and has an intrinsic molecular activity only 5% of that of DNA Polymerase I. This polymerase has 3'-5' exonuclease activity, is effective only on duplex DNA with gaps or single-strand ends of less than 100 nucleotides as template, and is inhibited by sulfhydryl reagents. EC 2.7.7.7.Multiplex Polymerase Chain Reaction: 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.Polymorphism, Restriction Fragment Length: 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.DNA Polymerase III: A DNA-dependent DNA polymerase characterized in E. coli and other lower organisms but may be present in higher organisms. Use also for a more complex form of DNA polymerase III designated as DNA polymerase III* or pol III* which is 15 times more active biologically than DNA polymerase I in the synthesis of DNA. This polymerase has both 3'-5' and 5'-3' exonuclease activities, is inhibited by sulfhydryl reagents, and has the same template-primer dependence as pol II. EC 2.7.7.7.Sequence Analysis, DNA: A multistage process that includes cloning, physical mapping, subcloning, determination of the DNA SEQUENCE, and information analysis.DNA, Protozoan: Deoxyribonucleic acid that makes up the genetic material of protozoa.Electrophoresis, Agar Gel: Electrophoresis in which agar or agarose gel is used as the diffusion medium.Gene Amplification: 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.DNA, Complementary: 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.Polymorphism, Genetic: 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.DNA Probes: 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.Alleles: Variant forms of the same gene, occupying the same locus on homologous CHROMOSOMES, and governing the variants in production of the same gene product.RNA, Viral: Ribonucleic acid that makes up the genetic material of viruses.Templates, Genetic: 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.RNA Polymerase I: 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.RNA Polymerase III: A DNA-dependent RNA polymerase present in bacterial, plant, and animal cells. It functions in the nucleoplasmic structure where it transcribes DNA into RNA. It has specific requirements for cations and salt and has shown an intermediate sensitivity to alpha-amanitin in comparison to RNA polymerase I and II. EC 2.7.7.6.Immunohistochemistry: Histochemical localization of immunoreactive substances using labeled antibodies as reagents.Immunoglobulin Heavy Chains: 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.Escherichia coli: A species of gram-negative, facultatively anaerobic, rod-shaped bacteria (GRAM-NEGATIVE FACULTATIVELY ANAEROBIC RODS) commonly found in the lower part of the intestine of warm-blooded animals. It is usually nonpathogenic, but some strains are known to produce DIARRHEA and pyogenic infections. Pathogenic strains (virotypes) are classified by their specific pathogenic mechanisms such as toxins (ENTEROTOXIGENIC ESCHERICHIA COLI), etc.RNA-Directed DNA Polymerase: 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.DNA Polymerase beta: A DNA repair enzyme that catalyzes DNA synthesis during base excision DNA repair. EC 2.7.7.7.Promoter Regions, Genetic: 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.Sequence Homology, Nucleic Acid: 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, Cultured: 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.Exons: 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.Restriction Mapping: Use of restriction endonucleases to analyze and generate a physical map of genomes, genes, or other segments of DNA.DNA, Neoplasm: DNA present in neoplastic tissue.Cell Line: Established cell cultures that have the potential to propagate indefinitely.Sequence Homology, Amino Acid: The degree of similarity between sequences of amino acids. This information is useful for the analyzing genetic relatedness of proteins and species.DNA Mutational Analysis: Biochemical identification of mutational changes in a nucleotide sequence.Nucleic Acid Hybridization: Widely used technique which exploits the ability of complementary sequences in single-stranded DNAs or RNAs to pair with each other to form a double helix. Hybridization can take place between two complimentary DNA sequences, between a single-stranded DNA and a complementary RNA, or between two RNA sequences. The technique is used to detect and isolate specific sequences, measure homology, or define other characteristics of one or both strands. (Kendrew, Encyclopedia of Molecular Biology, 1994, p503)In Situ Hybridization: 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.Gene Expression Regulation: 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.Phylogeny: The relationships of groups of organisms as reflected by their genetic makeup.RNA: 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)Sequence Alignment: 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.Cattle: 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.Species Specificity: The restriction of a characteristic behavior, anatomical structure or physical system, such as immune response; metabolic response, or gene or gene variant to the members of one species. It refers to that property which differentiates one species from another but it is also used for phylogenetic levels higher or lower than the species.Gene Expression Profiling: The determination of the pattern of genes expressed at the level of GENETIC TRANSCRIPTION, under specific circumstances or in a specific cell.Case-Control Studies: 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.Gene Frequency: The proportion of one particular in the total of all ALLELES for one genetic locus in a breeding POPULATION.Blotting, Western: 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.Time Factors: Elements of limited time intervals, contributing to particular results or situations.Point Mutation: 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.Oligodeoxyribonucleotides: 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.Gene Rearrangement: The ordered rearrangement of gene regions by DNA recombination such as that which occurs normally during development.Evaluation Studies as Topic: 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.Kinetics: The rate dynamics in chemical or physical systems.Immunoglobulin Light Chains: 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.Polymorphism, Single-Stranded Conformational: Variation in a population's DNA sequence that is detected by determining alterations in the conformation of denatured DNA fragments. Denatured DNA fragments are allowed to renature under conditions that prevent the formation of double-stranded DNA and allow secondary structure to form in single stranded fragments. These fragments are then run through polyacrylamide gels to detect variations in the secondary structure that is manifested as an alteration in migration through the gels.Nucleic Acid Amplification Techniques: Laboratory techniques that involve the in-vitro synthesis of many copies of DNA or RNA from one original template.Enzyme-Linked Immunosorbent Assay: An immunoassay utilizing an antibody labeled with an enzyme marker such as horseradish peroxidase. While either the enzyme or the antibody is bound to an immunosorbent substrate, they both retain their biologic activity; the change in enzyme activity as a result of the enzyme-antibody-antigen reaction is proportional to the concentration of the antigen and can be measured spectrophotometrically or with the naked eye. Many variations of the method have been developed.Paraffin Embedding: The infiltrating of tissue specimens with paraffin, as a supporting substance, to prepare for sectioning with a microtome.Feces: Excrement from the INTESTINES, containing unabsorbed solids, waste products, secretions, and BACTERIA of the DIGESTIVE SYSTEM.Blotting, Northern: 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.Recombinant Proteins: Proteins prepared by recombinant DNA technology.Transcription Factors: Endogenous substances, usually proteins, which are effective in the initiation, stimulation, or termination of the genetic transcription process.Phenotype: The outward appearance of the individual. It is the product of interactions between genes, and between the GENOTYPE and the environment.Tumor Virus Infections: 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, Neoplasm: RNA present in neoplastic tissue.Taq Polymerase: 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.-.DNA Replication: The process by which a DNA molecule is duplicated.Translocation, Genetic: A type of chromosome aberration characterized by CHROMOSOME BREAKAGE and transfer of the broken-off portion to another location, often to a different chromosome.RNA Replicase: 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)Papillomaviridae: 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.Dog Diseases: 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.Genetic Variation: Genotypic differences observed among individuals in a population.Reproducibility of Results: The statistical reproducibility of measurements (often in a clinical context), including the testing of instrumentation or techniques to obtain reproducible results. The concept includes reproducibility of physiological measurements, which may be used to develop rules to assess probability or prognosis, or response to a stimulus; reproducibility of occurrence of a condition; and reproducibility of experimental results.Oligonucleotide Array Sequence Analysis: 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.DNA-Binding Proteins: 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.Genes, Bacterial: The functional hereditary units of BACTERIA.Biopsy: Removal and pathologic examination of specimens in the form of small pieces of tissue from the living body.Electrophoresis, Polyacrylamide Gel: Electrophoresis in which a polyacrylamide gel is used as the diffusion medium.Binding Sites: The parts of a macromolecule that directly participate in its specific combination with another molecule.Tumor Cells, Cultured: 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.Bacterial Proteins: Proteins found in any species of bacterium.DNA, Fungal: Deoxyribonucleic acid that makes up the genetic material of fungi.Up-Regulation: 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 Diagnostic Techniques: MOLECULAR BIOLOGY techniques used in the diagnosis of disease.Oligonucleotides: 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)Plasmids: 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.Genetic Predisposition to Disease: A latent susceptibility to disease at the genetic level, which may be activated under certain conditions.Genetic Markers: A phenotypically recognizable genetic trait which can be used to identify a genetic locus, a linkage group, or a recombination event.Poly(ADP-ribose) Polymerases: 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.Viral Proteins: Proteins found in any species of virus.Protein Binding: 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.Cattle Diseases: Diseases of domestic cattle of the genus Bos. It includes diseases of cows, yaks, and zebus.Neoplasm Proteins: Proteins whose abnormal expression (gain or loss) are associated with the development, growth, or progression of NEOPLASMS. Some neoplasm proteins are tumor antigens (ANTIGENS, NEOPLASM), i.e. they induce an immune reaction to their tumor. Many neoplasm proteins have been characterized and are used as tumor markers (BIOMARKERS, TUMOR) when they are detectable in cells and body fluids as monitors for the presence or growth of tumors. Abnormal expression of ONCOGENE PROTEINS is involved in neoplastic transformation, whereas the loss of expression of TUMOR SUPPRESSOR PROTEINS is involved with the loss of growth control and progression of the neoplasm.Gene Expression Regulation, Neoplastic: Any of the processes by which nuclear, cytoplasmic, or intercellular factors influence the differential control of gene action in neoplastic tissue.Gene Rearrangement, B-Lymphocyte, Heavy Chain: 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.Swine: 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).DNA, Helminth: Deoxyribonucleic acid that makes up the genetic material of helminths.Herpesviridae Infections: Virus diseases caused by the HERPESVIRIDAE.Ligase Chain Reaction: A DNA amplification technique based upon the ligation of OLIGONUCLEOTIDE PROBES. The probes are designed to exactly match two adjacent sequences of a specific target DNA. The chain reaction is repeated in three steps in the presence of excess probe: (1) heat denaturation of double-stranded DNA, (2) annealing of probes to target DNA, and (3) joining of the probes by thermostable DNA ligase. After the reaction is repeated for 20-30 cycles the production of ligated probe is measured.Transfection: 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.Gene Library: 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.Neoplasm, Residual: Remnant of a tumor or cancer after primary, potentially curative therapy. (Dr. Daniel Masys, written communication)Gene Deletion: 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.Cell Line, Tumor: A cell line derived from cultured tumor cells.Pedigree: 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.Macromolecular Substances: 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.China: A country spanning from central Asia to the Pacific Ocean.Pregnancy: The status during which female mammals carry their developing young (EMBRYOS or FETUSES) in utero before birth, beginning from FERTILIZATION to BIRTH.RNA, Ribosomal, 16S: Constituent of 30S subunit prokaryotic ribosomes containing 1600 nucleotides and 21 proteins. 16S rRNA is involved in initiation of polypeptide synthesis.Chromosomes, Human, Pair 14: A specific pair of GROUP D CHROMOSOMES of the human chromosome classification.Cytomegalovirus: 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, Ribosomal: DNA sequences encoding RIBOSOMAL RNA and the segments of DNA separating the individual ribosomal RNA genes, referred to as RIBOSOMAL SPACER DNA.Flow Cytometry: 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, Immunoglobulin: 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).Heterozygote: An individual having different alleles at one or more loci regarding a specific character.Liver: A large lobed glandular organ in the abdomen of vertebrates that is responsible for detoxification, metabolism, synthesis and storage of various substances.Prevalence: 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.Myosin Heavy Chains: 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.Genes: 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.Protozoan Infections, Animal: Infections with unicellular organisms formerly members of the subkingdom Protozoa. The infections may be experimental or veterinary.Introns: 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.Asian Continental Ancestry Group: Individuals whose ancestral origins are in the southeastern and eastern areas of the Asian continent.Swine Diseases: Diseases of domestic swine and of the wild boar of the genus Sus.Codon: 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).Immunoenzyme Techniques: 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.Chromosome Mapping: Any method used for determining the location of and relative distances between genes on a chromosome.Chromosomes, Human, Pair 18: A specific pair of GROUP E CHROMOSOMES of the human chromosome classification.Models, Molecular: Models used experimentally or theoretically to study molecular shape, electronic properties, or interactions; includes analogous molecules, computer-generated graphics, and mechanical structures.Horse Diseases: Diseases of domestic and wild horses of the species Equus caballus.Tumor Markers, Biological: Molecular products metabolized and secreted by neoplastic tissue and characterized biochemically in cells or body fluids. They are indicators of tumor stage and grade as well as useful for monitoring responses to treatment and predicting recurrence. Many chemical groups are represented including hormones, antigens, amino and nucleic acids, enzymes, polyamines, and specific cell membrane proteins and lipids.Gene Rearrangement, gamma-Chain T-Cell Antigen Receptor: Ordered rearrangement of T-cell variable gene regions coding for the gamma-chain of antigen receptors.DNA Fingerprinting: 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.Molecular Weight: The sum of the weight of all the atoms in a molecule.Dogs: 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)Carrier Proteins: Transport proteins that carry specific substances in the blood or across cell membranes.Disease Outbreaks: Sudden increase in the incidence of a disease. The concept includes EPIDEMICS and PANDEMICS.Herpesvirus 6, Human: The type species of ROSEOLOVIRUS isolated from patients with AIDS and other LYMPHOPROLIFERATIVE DISORDERS. It infects and replicates in fresh and established lines of hematopoietic cells and cells of neural origin. It also appears to alter NK cell activity. HHV-6; (HBLV) antibodies are elevated in patients with AIDS, Sjogren's syndrome, sarcoidosis, chronic fatigue syndrome, and certain malignancies. HHV-6 is the cause of EXANTHEMA SUBITUM and has been implicated in encephalitis.Tissue Fixation: 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.Parvoviridae Infections: Virus infections caused by the PARVOVIRIDAE.Leukocytes, Mononuclear: 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.Papillomavirus Infections: Neoplasms of the skin and mucous membranes caused by papillomaviruses. They are usually benign but some have a high risk for malignant progression.Herpesvirus 4, Human: 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.Repetitive Sequences, Nucleic Acid: Sequences of DNA or RNA that occur in multiple copies. There are several types: INTERSPERSED REPETITIVE SEQUENCES are copies of transposable elements (DNA TRANSPOSABLE ELEMENTS or RETROELEMENTS) dispersed throughout the genome. TERMINAL REPEAT SEQUENCES flank both ends of another sequence, for example, the long terminal repeats (LTRs) on RETROVIRUSES. Variations may be direct repeats, those occurring in the same direction, or inverted repeats, those opposite to each other in direction. TANDEM REPEAT SEQUENCES are copies which lie adjacent to each other, direct or inverted (INVERTED REPEAT SEQUENCES).Fluorescent Antibody Technique, Direct: 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)Rats, Sprague-Dawley: 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.Infant, Newborn: An infant during the first month after birth.Cytomegalovirus Infections: 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.Polymorphism, Single Nucleotide: A single nucleotide variation in a genetic sequence that occurs at appreciable frequency in the population.Chlamydia Infections: Infections with bacteria of the genus CHLAMYDIA.Sequence Deletion: Deletion of sequences of nucleic acids from the genetic material of an individual.Disease Models, Animal: 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.Clone Cells: 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)Membrane Proteins: 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.Homozygote: An individual in which both alleles at a given locus are identical.HeLa Cells: 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.Specimen Handling: Procedures for collecting, preserving, and transporting of specimens sufficiently stable to provide accurate and precise results suitable for clinical interpretation.Bone Marrow: 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.Aborted Fetus: A mammalian fetus expelled by INDUCED ABORTION or SPONTANEOUS ABORTION.RNA, Ribosomal, 18S: Constituent of the 40S subunit of eukaryotic ribosomes. 18S rRNA is involved in the initiation of polypeptide synthesis in eukaryotes.Herpesviridae: 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, Kinetoplast: DNA of kinetoplasts which are specialized MITOCHONDRIA of trypanosomes and related parasitic protozoa within the order KINETOPLASTIDA. Kinetoplast DNA consists of a complex network of numerous catenated rings of two classes; the first being a large number of small DNA duplex rings, called minicircles, approximately 2000 base pairs in length, and the second being several dozen much larger rings, called maxicircles, approximately 37 kb in length.Gene Expression Regulation, Enzymologic: Any of the processes by which nuclear, cytoplasmic, or intercellular factors influence the differential control of gene action in enzyme synthesis.In Situ Hybridization, Fluorescence: 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.Prognosis: 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.Saccharomyces cerevisiae: 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.BrazilFluorescent Antibody Technique, Indirect: A form of fluorescent antibody technique commonly used to detect serum antibodies and immune complexes in tissues and microorganisms in specimens from patients with infectious diseases. The technique involves formation of an antigen-antibody complex which is labeled with fluorescein-conjugated anti-immunoglobulin antibody. (From Bennington, Saunders Dictionary & Encyclopedia of Laboratory Medicine and Technology, 1984)Predictive Value of Tests: In screening and diagnostic tests, the probability that a person with a positive test is a true positive (i.e., has the disease), is referred to as the predictive value of a positive test; whereas, the predictive value of a negative test is the probability that the person with a negative test does not have the disease. Predictive value is related to the sensitivity and specificity of the test.Down-Regulation: 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.Prospective Studies: 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.Genome, Viral: The complete genetic complement contained in a DNA or RNA molecule in a virus.Bacteriophage T7: 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.Chlamydia trachomatis: Type species of CHLAMYDIA causing a variety of ocular and urogenital diseases.DNA Methylation: Addition of methyl groups to DNA. DNA methyltransferases (DNA methylases) perform this reaction using S-ADENOSYLMETHIONINE as the methyl group donor.Mycoplasma Infections: Infections with species of the genus MYCOPLASMA.Cat Diseases: 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.Bacteriological Techniques: Techniques used in studying bacteria.Digoxigenin: 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.Genes, ras: 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.Temperature: 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.Mice, Inbred C57BLFatal Outcome: 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.Microsatellite Repeats: 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).

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;
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.
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.
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 ...
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, ...
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.
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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 ...
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 ...
Performance of MiniPCRTM mini8, a portable thermal cycler - MiniPCRTM mini8 Thermal Cycler;GeneAmp®PCR system 9700;performance;STR multiplex kits;mtDNA HV1/HV2;
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 ...
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 ...
Watch this video to learn how researchers at Sangamo BioSciences use Bio-Rads QX100 Droplet Digital PCR system to quantify the level of HIV reservoir in treatment subjects with their unique zinc-finger-based therapy. At approximately one copy of HIV in 1,000 cells analyzed, this quantification would present a challenge for conventional methods. The Droplet Digital PCR system allows for the quantification of target DNA with unrivaled precision by partitioning samples into 20,000 nanoliter-sized droplets. After PCR on a thermal cycler, PCR-positive and PCR-negative droplets are counted to provide absolute quantification of the target DNA. The QX100 Droplet Digital PCR system is the third generation of PCR technology and provides an absolute measure of target DNA molecules with unrivaled accuracy, precision, and sensitivity.
AccuPrep® GMO DNA Extraction Kit allows the extraction of DNA from agricultural products such as beans, corn, and rice or from processed foods such as bean sprouts, tofu and condensed milk, for the detection of GMOs. The kit includes all reagents required for extraction and in addition uses a column type extraction system to allow a more rapid, more convenient extraction of higher-quality DNA when compared to conventional methods. A high amount of high-purity DNA can be extracted; for example, 20 ~ 40 ug of DNA for 1 g of powdered bean, with an A260/A280 ratio of over ...
The synthetic gox gene. An open reading frame (ORF; EMBL Bank: GU214711.1) of 1296 bp was previously reported as a goxgenesequence. The sequence was optimized based on the canola plant codon preference, and the regulatory Kozak sequence was added before the start codon to improve the efficiency of transcription and translation (Kozak, 1989; Gustafsson et al. 2004). The BamHI and SacI restriction sites were introduced at the 5 and 3 ends of the synthetic gene, respectively. The synth-gox gene was synthesized by Shine Gene Molecular Biotech, Incorporated (Shanghai, China), and the sequence was submitted to GenBank (Accession Number, HQ110097).. Construction of plant expression vector. The synth-gox gene was used to replace the gus fragment in the binary vector pBI121 (Clontech) using BamHI and SacI restriction sites. By this strategy, the synth-gox gene came under the control of the constitutive cauliflower mosaic virus (CaMV) 35S promoter and nopaline synthase (nos) terminator (pBI-synth-gox) ...
Two different molecular approaches, a multiplex PCR and PCR-RFLP of ITS-rDNA, were developed for the identification of Hoplolaimus species. DNA sequences of H. columbus, H. galeatus, H. concaudajuvencus, H. magnistylus, H. seinhorsti and three undescribed Hoplolaimus species were used to design species-specific primers. Three reverse species-specific PCR primers for H. columbus, H. galeatus and H. magnistylus were developed using the ITS1 region exhibiting interspecific variation. Three species-specific PCR primers in combination with the forward primer, Hoc-1f, produced distinct amplicons of 580 bp for H. columbus, 120 bp for H. galeatus and 340 bp for H. magnistylus. We successfully identified each of three species by multiplex PCR when all three were mixed in a single PCR reaction. Restriction enzyme digests of the PCR amplicon using HaeIII and RsaI permitted discrimination of H. columbus, H. galeatus, H. magnistylus, H. concaudajuvencus, H. sp. 1, H. sp. 2 and H. sp. 3 from each other. These results
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Polymerase chain reaction Polymerase chain reaction (PCR) is a molecular biology technique[1] for enzymatically replicating DNA without using a living
We have developed teabags packed with dehydrated plant powders, without any supplements, for preparation of plant infusions necessary to develop media for culturing rhizobacteria. These bacteria are efficiently cultivated on such plant teabag culture media, with better progressive in situ recoverability compared to standard chemically synthetic culture media. Combining various plant-based culture media and incubation conditions enabled us to resolve unique denaturing gradient gel electrophoresis (DGGE) bands that were not resolved by tested standard culture media. Based on polymerase chain reaction PCR-DGGE of 16S rDNA fingerprints and sequencing, the plant teabag culture media supported higher diversity and significant increases in the richness of endo-rhizobacteria, namely Gammaproteobacteria (Enterobacteriaceae) and predominantly Alphaproteobacteria (Rhizobiaceae). This culminated in greater retrieval of the rhizobacteria taxa associated with the plant roots. We conclude that the plant teabag ...
On Tuesday, We did agarose gel electrophoresis for PCR samples to check for amplification 10:08, 23 October 2012. we did not get any amplification in DNA samples however, we got the positive control band. We made an interpretation from the gel that may be primers did not bind at their specific sites or may be there is some problem in DNA sample. Since we are running out of our DNA sample, we decided to do the DNA extraction.This time we will be comparing two different protocols on parallel. One that we used before i.e, DNA extraction for E.coli chromosome and other that group 25 used which is, Bacterial Genomic DNA extraction protocol. On Wednesday, we did the culture to grow the E. coli. cells in four flasks and incubated in shaker for overnight. On Thursday, since we did not have enough killing buffer we did the extraction with the genomic DNA extraction protocol only. We put rest of the three culture flasks at 4ºC.We will do agarose gel electrophoresis for these DNA samples in our next lab. ...
Blood samples were collected at the study sites, processed to plasma aliquots, and sent to the central laboratory for quantitative CMV DNA polymerase chain reaction (PCR) testing. Plasma samples were assayed for CMV concentration using a qualified PCR method. This method was linear over 200-100,000 viral copies/mL with a lower limit of quantification (LLOQ) of 200 copies/mL. Results below LLOQ were considered undetectable. Confirmed undetectable plasma CMV DNA within 6 weeks was defined as 2 consecutive post-baseline, on-treatment undetectable results separated by ,/= 5 days (assessed by the central laboratory). Samples were collected on Days 1 and 8, weekly during Weeks 2-6, and once in Weeks 8, 10, 12, 16, 20, 24 (treatment) and Weeks 1, 4, 8, 12 (follow-up). Permissible assessment windows were: Days 8-15 +/- 1 day; Weeks 3-4 +/- 2 days; Weeks 5-6 +/- 3 days; Weeks 8-12 +/- 4 days; Weeks 16-24 +/- 7 days (treatment) and Weeks 1-4 +/- 2 days; Weeks 8-12 +/- 4 days (follow-up ...
Autor: Schutze, T. et al.; Genre: Zeitschriftenartikel; Im Druck veröffentlicht: 2011; Keywords: Chemical Fractionation/*methods; DNA/*genetics/*isolation & purification; Emulsions; Humans; Oils/*chemistry; Polymerase Chain Reaction/*methods; Time Factors; Water/*chemistry; Titel: A streamlined protocol for emulsion polymerase chain reaction and subsequent purification
Abstract: For disease control and eradication and in countries trying to establish a disease free status, effective diagnostic is a paramount. Many diagnostics find its application throughout different levels of laboratory research processes. Polymerase Chain Reaction (PCR) is one of the most important molecular diagnostic tools which allow the detection of nucleic acid targets. Because of its excellent sensitivity, specificity and speed, PCR has rapidly become the widely used molecular biological techniques in scientific, medical and research fields. There are different types of PCRs which are used specifically for certain specific purposes. The types of PCR and their applications are discussed in this review article. ...
www.MOLUNA.de Single Cell Diagnostics [4221608] - This book applies modern molecular diagnostic techniques to the analysis of single cells, small numbers of cells, or cell extracts. 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
There is a long and growing list of gene translocation events that are linked to cancer. Whether the result of intra- or interchromosomal exchanges, these translocations commonly involve genes encoding a kinase or a transcription factor. The resulting fusion genes are often the principal drivers of tumorigenesis and therefore serve as diagnostic markers and/or targets for specific therapies (e.g., kinase inhibitors). Fusion mRNAs from translocation events can be detected by highly sensitive methodologies based on polymerase chain reaction (PCR); however, these approaches can be frustrated by the fact that a particular target gene may be fused to any of more than a dozen different partner genes, requiring numerous primers to cover all possible fusion events. In contrast, FISH can detect a translocation-related break in a target gene irrespective of which partner gene has been fused to it. This is done by labeling two pools of probes with different fluorophores; for example, one pool may be ...
Read this full essay on animal genomic dna extraction. For the 2C experiment, we have been using chickens muscle tissue sample to obtain the DNA sample. The...
The qBiomarker Somatic Mutation PCR Assays are a fast and reliable mutation analysis tool using ARMS® (Amplification Refractory Mutation System) primer-based allele discrimination and Hydrolysis Probe-based quantitative real-time PCR technology. It utilizes a proprietary and experimentally verified algorithm for designing mutation-specific qPCR primers and probe. Each qBiomarker Somatic Mutation PCR Assay is subjected to rigorous experimental verification. Both sensitive detection of the intended mutation in a sample (as low as 1% mutant sample on a wild-type sample background) and assay specificity (i.e. discrimination between mutant sample and wild-type sample) are guaranteed when used with the qBiomarker Probe Master Mix. A gene-specific reference assay for checking sample quality and measuring sample input and gene copy dosage is included for each mutation assay. See the qBiomarker Somatic Mutation PCR Array System User Manual/ Handbook for protocol to follow.. The qBiomarker Somatic ...
The qBiomarker Somatic Mutation PCR Assays are a fast and reliable mutation analysis tool using ARMS® (Amplification Refractory Mutation System) primer-based allele discrimination and Hydrolysis Probe-based quantitative real-time PCR technology. It utilizes a proprietary and experimentally verified algorithm for designing mutation-specific qPCR primers and probe. Each qBiomarker Somatic Mutation PCR Assay is subjected to rigorous experimental verification. Both sensitive detection of the intended mutation in a sample (as low as 1% mutant sample on a wild-type sample background) and assay specificity (i.e. discrimination between mutant sample and wild-type sample) are guaranteed when used with the qBiomarker Probe Master Mix. A gene-specific reference assay for checking sample quality and measuring sample input and gene copy dosage is included for each mutation assay. See the qBiomarker Somatic Mutation PCR Array System User Manual/ Handbook for protocol to follow.. The qBiomarker Somatic ...
A stable, latent reservoir for HIV-1 in resting CD4+ T cells is "the principle barrier to a cure," the researchers say. Quantitative outgrowth assays and assays for cells that produce viral RNA after T-cell activation may underestimate the reservoir size because 1 round of activation does not induce all proviruses. Many studies, the researchers say, rely on simple assays based on polymerase chain reaction to detect proviral DNA regardless of transcriptional status, but the clinical relevance of those assays is unclear since the vast majority of proviruses are defective. In their study, supported by the National Institute of Allergy and Infectious Diseases, the researchers analyzed DNA sequences from , 400 HIV proviruses from 28 people with HIV. They mapped 2 types of flaws: deletions and lethal mutations. They then developed strategically placed "genetic probes" that could distinguish between deleted or highly mutated proviruses and intact ones. Finally, they developed a nanotechnology-based ...
The affordability, utility, and possibilities afforded by genetic "barcodes" has captured the interest of researchers, agencies, and funders on an international level. This has led to an explosion of genetic data generation and public availability of digital DNA on barcode databases such as NCBI Genbank and BOLD (Barcode of Life database). 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. The use of qPCR as a means of species detection by fisheries biologists has risen over the past decade. As a result, the adoption of qPCR has led to the transition of the qPCR technique from being purely a laboratory technique to a field technique. This transition requires much scrutiny, development, and adaptation to become wholly accepted by researchers and resource managers in the fisheries biology community. In the past 3 years, in ...
Researchers determined which cytokines induced the two activated antimicrobial macrophage effector reactions-resistance to infection and intracellular killing-and which cytokines shut down the killing. The author made inroads in the understanding of Francisella infections, devised a polymerase chain reaction-based detection system and an attenuated and subunit vaccine, and explored the use of immunomodulation with bacille-Calmette-Guerin (BCG) with great success. In this chapter the author seeks additional philanthropic funds to assist companies and academic investigators in the clinical development of new anti-TB drugs. The path is set for the foundation, and there is great excitement both within the foundation and the research community. Sequella, Inc., is working with investigators in universities around the world and has licensed some innovative, useful products for diagnosis and treatment of tuberculosis (TB). It is currently raising money to fund the development of its products and hopes to have
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Fungi play important roles in the ecosystem processes such as nutrient cycling and degradation. However, the majority of fungi are unknown and difficult to isolate and culture. Due to the limitations of culture-based methods and traditional morphological observation, fungal ecologists turn to utilize molecular techniques such as nucleic acid hybridization,DNA sequencing,DNA fingerprinting techniques for the studies of species identification,population structure and community diversity. The present paper summarizes the theories of these molecular techniques and their applications in the study of fungal ecology.
2015 Sexually transmitted diseases treatment guidelines. Centers for Disease Control and Prevention website. Available at: http://www.cdc.gov/std/tg2015/default.htm. Updated March 9, 2016. Accessed June 6, 2016.. Beauman JG. Genital herpes: a review. Am Fam Physician. 2005;72(8):1527-1534.. Gardella C, Huang ML, Wald A, et al. Rapid polymerase chain reaction assay to detect herpes simplex virus in the genital tract of women in labor. Obstet Gynecol. 2010;115(6):1209-1216.. Genital herpes-CDC fact sheet (detailed). Centers for Disease Control and Prevention website. Available at: http://www.cdc.gov/std/herpes/stdfact-herpes-detailed.htm. Updated November 17, 2015. Accessed June 6, 2016.. ...
Background. PCR amplicon sequencing has been widely used as a targeted approach for both DNA and RNA sequence analysis. Highly multiplex PCR has further enabled the enrichment of hundreds of amplicons in one simple reaction. At the same time, the performance of PCR amplicon sequencing can be negatively affected by issues such as high duplicate reads, polymerase artifacts and PCR amplification bias. Recently, people have made good progress at addressing those shortcomings by incorporating molecular barcodes into PCR primer design. So far, most work has been demonstrated using one to a few pairs of primers, which limits the size of the region one can analyze at one time.. Results. We described a simple protocol, which enables the use of molecular barcodes in highly multiplex PCR with hundreds of amplicons. Using this protocol and reference materials, we studied how molecular barcodes can increase the accuracy of variant calling at very low allelic frequency and reduce PCR amplification bias. We ...
The PCR replicated the wanted DNA fragments from the patient. the PCR will heat up to 95 degree Celsius and cool down to 50 degree Celsius and then heat back up to 72 degree Celsius within one cycle. Over all there will be 30 cycles. At the end of the 30 cycles we have over a billion of the wanted fragments and 60 unwanted DNA molecule strands in the solution. Step by Step instruction to amplify the Patients DNA Sample: 1. Need to extract the DNA from the patient. 2. Put the DNA into a special PCR tube. 3. Add primer #1 to the PCR tube with the DNA. 4. Add primer #2 to the PCR tube with DNA. 5. Add Nucleotides (the A,C,T,and G). 6. Add the DNA polymerase to the PCR tube. 7. Place the PCR tube into the thermal cycler. 8. Set the temperature of the thermal cycler to 95 degree Celsius and set the machine to run 30 cycles. 9. Now the thermal cycler cools down to 50 degree Celsius and primer #1 and #2 attach to the single strands of DNA. 10. Now the thermal cycler temperature changes to 72 degree ...
Note: Optimal annealing temperatures are often higher than the Tm of the primers (approximately +5 to +10°C) and have to be determined empirically. For both primers, the Tm should be similar.. Bases that do not hybridize to the template may be added at the 5´ end of a primer (e.g., for introducing restriction sites into the amplification product). Primer concentrations between 0.1 and 0.6 µM are generally optimal. Higher primer concentrations may promote mispriming and accumulation of nonspecific product. Lower primer concentrations may be exhausted before the reaction is completed, resulting in lower yields of desired product.. Note: For some systems, a higher primer concentration (up to 1 µM) may improve results. When testing new primers, always include a positive control reaction with a template that has been tested for function in PCR. This control shows whether the primers are working. The Human Genomic DNA from Roche is a good control template for evaluation of human primer ...
Pjevac, P.; Schauberger, C.; Poghosyan, L.; Herbold, C.W.; Kessel, M.A.H.J. van; Daebeler, A.; Steinberger, M.; Jetten, M.S.M. ; Lücker, S.; Wagner, M.; Daims, H. ...
GenemerTM Products. The GenemerTM product line is PCR based. The product includes a specific primer pair for gene or mutation specific amplification. Genemer products are available for the gene fragment and disorders listed. Specialized optimized conditions may be required for certain triple repeat disorder amplifications. The GenemerTM kit is a complete easy-to-use kit for reliable genotyping of a gene fragment. The product includes a specific primer pair for gene or mutation specific amplification, optimized buffers and dNTPs and in most cases, control DNA. These kits contain specialized and optimized conditions that are required for amplification of large repeats in certain triple repeat disorder amplifications. Gene Link recommends these GenemerTM kits for researchers who have not established their own optimized amplification conditions. GenemerTM kits are also available for conventional radioactive-based detection methods. A Radioactive component is not present in these kits. Gene Link ...
Creative Biogene HSV Ⅰ&Ⅱ Typing real time PCR kit is used for the detection of HSV genotypeⅠ & genotype Ⅱ in serum, herpes secretion or genital swabs samples by using real time PCR systems. The kit contains a specific ready-to-use system for the detection of the HSVgenotypeⅠ& genotype Ⅱ by polymerase chain reaction in the real-time PCR system. The master contains reagents and enzymes for the specific amplification of the HSV genotypeⅠ & genotype Ⅱ DNA. Fluorescence is emitted and measured by the real time systems optical unit during PCR.http://www.creative-biogene.com/HSV-%E2%85%A0-%E2%85%A1-Typing-Real-Time-PCR-Kit-PDHS-DR003-1290583-88.html. ...
gapdh is another common sequence that is used as a ubiquitous signal but I dont know if its applicable for you a.boasso wrote: , We are using a semi-quantitative pcr assay to evaluate the expression level , of genes involved in the immune system functions (like citokynes, receptors, , co-stimulation molecules), in hiv+ individuals or aids patients. , In this assay we perform a pcr reaction with two sets of primers: , 1. a pair of primers for the target mRNA amplification (citokynes, receptors , or co-stimulation molecules) , 2. a pair of prime for the beta-actin mRNA, which will be the internal , standard , , The beta-actin mRNA concentration is determined in a previous step, with a , specific competitive pcr assay, with a synthetic competitor, used in , different reactions at different concentrations. , , Some critics have been moved against the use of beta-actin as an internal , standard, as its expression seems to change widely in pathologic cells. , , Is anybody able to give me some hints ...
This compact personal thermal cycler is ideal for the classroom! Utilize our thermal gradient technology for optimal PCR. Educational discount available.
The gSYNC™ DNA Extraction Kit is a genomic DNA extraction kit optimized for genomic, mitochondrial and virus DNA purification from whole blood (fresh blood and frozen blood), serum, plasma, buffy coat, body fluids, cultured cells, tissue, rodent tails, ear punches, formalin-fixed paraffin-embedded tissue (FFPE), amniotic fl
The quantitative polymerase chain reaction (qPCR) is a relatively new technique that combines the reliabiity of regular PCR with real-time screening.
In this months Alkami PCR Reviews Dr. Michael Zwick discusses the Advantages and Disadvantages of PCR-Based Detection Kits. This month I conclude my three part series on why I feel PCR-based diagnostics should be common in the clinical lab setting. Last month, I discussed three reasons why PCR is particularly useful, i.e., that it is fast, doesnt require culturing, and is cost-effective. Today I will discuss some additional rationale for considering the use of PCR in the clinical setting.. You can find his article at http://www.alkami.com/reviews/mszver03.htm The Alkami Biosystems site, http://www.alkami.com, is a comprehensive resource for the polymerase chain reaction (PCR). We have online references that include primer design tips and tools, reagent concentrations, polymerase characteristics, PCR methods, and temperature calculations.In addition, you will find regular reviews of PCR issues, employment resources for the biologist, as well as special FREE lab supply offers at ...
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Polymerase Chain Reaction (PCR) Products/Tools Report Detail: With a CAGR of 10.1%, global market value for PCR Products/Tools market is anticipated to be worth US$12 billion by 2020. PCR Reagents and PCR Detection Kits/Assays accounts for more than 40% of the market share and fastest growing segment with a CAGR approximately 10.8% and 10.5% by…
Relative fluorescent quantitation (or quantitative fluorescence PCR (QF-PCR)) is a technique used in a variety of fragment analysis applications that requires accurate peak height comparisons across multiple samples. Applications that utilize this t
DI-fusion, le Dépôt institutionnel numérique de lULB, est loutil de référencementde la production scientifique de lULB.Linterface de recherche DI-fusion permet de consulter les publications des chercheurs de lULB et les thèses qui y ont été défendues.
The pilot study was performed using two genetically homogeneous Punjabi cohorts, one resident in the United Kingdom and one indigenous to Pakistan. Subjects with (N = 1732) and without (N = 1780) type 2 diabetes were genotyped for thirteen circadian variants using a competitive allele-specific polymerase chain reaction method. Associations between the SNPs and type 2 diabetes were investigated using logistic regression. The results were also combined with in silico data from other South Asian datasets (SAT2D consortium) and white European cohorts (DIAGRAM+) using meta-analysis. The rs7602358G allele near PER2 was negatively associated with type 2 diabetes in our Punjabi cohorts (combined odds ratio [OR] = 0.75 [0.66-0.86], p = 3.18×10−5), while the BMAL1 rs11022775T allele was associated with an increased risk of the disease (combined OR = 1.22 [1.07-1.39], p = 0.003). Neither of these associations was replicated in the SAT2D or DIAGRAM+ datasets, however. Meta-analysis of all the cohorts ...
Real-time PCR has become a standard tool for analyzing mRNA expression levels, e.g., for validation of data obtained from whole-genome approaches (e.g., microarrays). For data interpretation, however, real-time PCR instrument software mainly focuses on raw data and does not support analysis of multiple runs or plates. Software tools that improve and accelerate the exploitation and statistical analysis of extended studies are therefore needed.. We studied the expression of 16 transcripts that might be predictive for the progression stage of spinocerebellar ataxia 1 and 3 (SCA1 + 3), two autosomal-dominant neurodegenerative diseases. These transcripts had previously been selected based on microarray experiments because of aberrant expression patterns in comparison to reference genes. The LightCycler® 480 Multiple Plate Analysis Software was tested as a novel tool for the compiled analysis of multiple LightCycler® 480 Instrument runs performed during this study.. Back to Top. ...
Using this strategy, we calculated log IC50 values from the resulting inhibition curves. Greater than a half log increase in IC50 was observed for both the IE and gB targets of ddPCR over qPCR for both SDS (absolute log difference in IC50 qPCR vs ddPCR IE, 0.554, and vs ddPCR gB, 0.628) and heparin (absolute log difference in IC50 qPCR vs ddPCR IE, 0.655, and vs ddPCR gB, 0.855). The probability of difference between the data sets for ddPCR and qPCR was ,99.99% for both inhibitors and both ddPCR targets, indicating that ddPCR tolerated the presence of these inhibitors better than qPCR. However, this difference was not noted when we compared ddPCR and qPCR in the presence of EDTA for both ddPCR targets (log difference in IC50 qPCR vs ddPCR IE, 0.116, and vs ddPCR gB, 0.0198), possibly owing to different inhibition mechanisms. EDTA is a calcium chelator, whereas SDS and heparin both act on DNA polymerase.. The ddPCR CMV assay is more tolerant to SDS and heparin than the qPCR assay, indicating that ...
A process for analyzing length polymorphism in DNA regions wherein the following steps are carried out: (a) annealing at least one primer pair to the DNA to be analyzed, wherein one of the molecules of the primer pair is substantially complementary to one of the complementary strands of the 5 or 3 flank of a simple or cryptically simple DNA sequence, and wherein the annealing occurs in such an orientation that the synthesis products obtained by a primer-controlled polymerisation reaction with one of said primers can serve as template for annealing the other primer after denaturation; (b) primer-controlled polymerase chain reaction; and (c) separating and analyzing the polymerase chain reaction products.
PubMed comprises more than 30 million citations for biomedical literature from MEDLINE, life science journals, and online books. Citations may include links to full-text content from PubMed Central and publisher web sites.
Reverse transcription quantitative polymerase chain reaction analysis of genes in K562 cells treated with virosecurinine at 6.25, 25 and 50 μmol/l for 48 h. mT
In situations where the amount of available sample DNA is limited, or where there is a low level of pathogen DNA mixed with a high level of host DNA, and we wish to identify the pathogen, it can be helpful to amplify the target organism by PCR. For bacterial species identification, the 16S ribosomal RNA gene can be amplified from all bacteria non-specifically, without amplifying eukaryotic host DNA, or viruses. As with the whole-genome species ID approach shown in Fig. 1, we have found bead-beating to lyse cells rapidly, yielding DNA with a sufficiently high fragment length for amplification of the 1.5 kb 16S gene. There is no need to purify the extracted DNA before PCR. Fast polymerases are available which can process 1 kb of template in around 30 seconds, meaning that 16S PCR can be performed on a standard thermocycler in 25 minutes. If PCR is performed using Oxford Nanopores modified primers, sequencing adapters can be attached rapidly following amplification, by chemical ligation. This ...
The polymerase chain reaction (PCR) is a laboratory technique for DNA replication that allows a target DNA sequence to be selectively amplified. PCR can
Reverse transcription polymerase chain reaction (RT‐PCR) expression patterns of the genes coding for Laccaria bicolor Zn finger protein and Ser/Threo protein
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Scientist placing test tubes in thermocycler used for polymerase chain reaction in laboratory. Filmed in Oxford, England, UK. - Stock Video Clip K002/8129
There are currently various different tests for HIV infections such as HIV antibody test, P24 antigen test, Polymerase chain reaction test (PCR), Fourth generation test a..
The PCR (template) DNA must be highly purified DNA having 30ng to 50ng concentration, 50% to 55% GC content and free from chemical contaminants and other DNA contaminants.
What is the purpose of displayPROFILETM-RFDD? DisplayPROFILETM RFDD is a method which uses the polymerase chain reaction (PCR) to make consistent and rapid gene expression profiles and to identify differentially expressed genes in various tissues and cells. To produce molecular snapshots of expressed genes, the displayPROFILETM technique involves digesting double-stranded cDNA with a restriction enzyme, ligating specific DNA adaptors to the cDNA fragments, and annealating specific primers onto the adaptors (Display Sytems Biotech). The cDNA fragments are then divided into 64 different PCR reactions. Each PCR reaction amplifies a different expression window, or a set of 400 or more cDNA fragments. Unlike standard differential display systems that use poly-A primed PCR amplification, RFDD-PCR uses the TaqI restriction enzyme and specific PCR primers for amplification (Display Systems Biotech). As a result of not relying on poly-A-primed PCR, this technique can be used for both eukaryotic and ...
The use of magnetic particles in many fields of biochemistry, molecular biology, and medicine has been well documented and several magnetic particles are now available for diagnostic and cell...
View Notes - Biol110-10-Lecture 2-Methods from BIOL 110 at UCSC. How are cells and their components studied? Cell biological techniques: Microscopy Biochemistry Molecular Biology Genetics Assigned
The Ct or threshold cycle value is the cycle number at which the fluorescence generated within a reaction crosses the fluorescence threshold, a fluorescent signal significantly above the background fluorescence. At the threshold cycle, a detectable amount of amplicon product has been generated during the early exponential phase of the reaction. The threshold cycle is inversely proportional to the original relative expression level of the gene of interest ...
The LA PCR Genome Set includes highly purified, high molecular weight human genomic DNA PCR controls, E.coli genomic DNA PCR controls and appropriate primers. These templates and primers are intended for use as long range PCR controls and provide a useful method for optimizing PCR conditions given a wide variety of templates.. ...
suitable cell lysis buffer - posted in Tissue and Cell Culture: Hi I need to do some elisa on my CHO cell line, is there any suitable cell lysis buffer that I can prepare in lab? regards
First-strand cDNA Synthesis System for Quantitative RT-PCR has been designed for the highest efficiency conversion of RNA to cDNA and is fully optimized for quantitative real-time PCR applications.
Bacteria is a prokaryotic organism having a large number of species. They are present in most of the habitats of earth. Some bacterias are useful while others are pathogenic and even threatful to life. Knowledge of the true bacterial species is fundamental for the effective utilization of that particular species . Most of the bacterias have not been characterized and only some of the species can be grown in labortary. Traditional methods of bacterial identification rely on phenotypic identification using gram staining, culture and biochemical methods. However, these methods of bacterial identification suffer from some drawbacks. The identification and characterization of bacterial species associated with particular traits can be done by using molecular traits.The most common way of molecular characterization of bacteria is by using Polymerase Chain Reaction. This technology allows us to identify any bacterial species to any level of precision by using a single cell. The technology uses molecular ...
The availability of simple methods for purification of DNA and RNA has greatly facilitated the analysis and characterization of the genome and gene expression. There is a demand to isolate DNA and RNA rapidly and conveniently from a variety of cellular sources, including cells and tissues from mammalian, plant and bacterial cultures.
ddPCR™ probe assay designed for gene expression analysis. Probe assays consist of unlabeled PCR primers and a dual labeled fluorescent probe. Info: HEX; Same primer pair and probe as used in qPCR assay qRnoCIP0037965; intron_spanning ...
Research in the Department of Biochemistry encompasses very diverse questions and uses a wide variety of approaches, experimental systems, and techniques. Nevertheless, what bonds us is an interest in understanding fundamental biological questions at the level of how molecules act and interact to accomplish highly complex, intra- and intercellular processes. Our diversity enriches our intellectual environment and provides an incredibly broad spectrum of expertise that benefits all of us, as we tackle a wide variety of important questions. All of us study molecules: proteins, RNA, DNA, and polyphosphate; we analyze their synthesis, structure, actions and interactions. We use physical techniques such as spectroscopy, laser light traps and crystallography, cell biological techniques such as light microscopy and cell fractionation, biochemical techniques such as enzyme purification and characterization, along with molecular biological techniques and genetics. By attacking problems using these ...
TransScript® First-Strand cDNA Synthesis SuperMix,RT-PCR,PCR, RT-PCR, qPCR, qRT-PCR,Products,Beijing TransGen Biotech Co.Ltd,OverviewContents& storageCitations & referencesRelated ImagesDownloadOverviewDescription&bull TransScript First-Strand
Polymerase chain reaction (PCR) enables researchers to produce millions of copies of a specific DNA sequence in approximately two hours. This automated process bypasses the need to use bacteria for amplifying DNA.
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 ...
Cytomegalovirus DNA was detected in 18 of 19 eyes with untreated cytomegalovirus retinitis. We detected cytomegalovirus DNA in 19 of 40 vitreous samples from patients with previously treated cytomegalovirus retinitis. Cytomegalovirus DNA was not detected in any of 69 patients who did not have a clinical diagnosis of cytomegalovirus retinitis. Thus, the assay had an estimated sensitivity of 95% in detecting untreated cytomegalovirus retinitis and a sensitivity of 48% in detecting cytomegalovirus retinitis that had been treated with systemic ganciclovir or foscarnet, or both. The assay did not give false-positive results in patients with vitreous hemorrhage or vitreous inflammation. Most important, the assay did not give false-positive results in AIDS patients with vitreous inflammation from causes other than cytomegalovirus retinitis.. ...
Acetyl-coenzyme A carboxylase α (ACC-alpha) is considered as the key regulatory enzyme in fatty acid biosynthesis. ACC-alpha gene is located on Caprine chromosome 11 and is polymorphic in many goat breeds. In the current study, we aimed to find possible single nucleotide polymorphisms (SNPs) in the exon 1 region of the ACC-alpha gene in Iranian Mahabadi goat breed. Genomic DNA was extracted from blood samples of 150 Mahabadi does. The exon 1 region of the ACC-alpha gene was amplified to produce a 390 bp fragment. The PCR products were analyzed by both polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) techniques. RFLP was performed utilizing HinfI endonuclease enzyme. No polymorphism was observed after digestion of the PCR products using HinfI. However, SSCP of the PCR products revealed two conformation patterns at the exon 1 region of goat ACC-alpha gene with frequencies of 86% and 14%,
TY - JOUR. T1 - Cryptosporidium parvum mixed genotypes detected by PCR-restriction fragment length polymorphism analysis. AU - Reed, C.. AU - Sturbaum, G. D.. AU - Hoover, P. J.. AU - Sterling, Charles R. PY - 2002. Y1 - 2002. N2 - Combinations of 10 Cryptosporidium parvum oocysts, with various ratios of genotype I to genotype II, were isolated and subjected to PCR-restriction fragment length polymorphism analysis. Amplification of both genotypes in these samples ranged from 31 to 74% and yielded no information about the genotype proportions. In addition, since both genotypes were not always detected, amplification of a single genotype is not conclusive evidence that the sample contains only a single genotype.. AB - Combinations of 10 Cryptosporidium parvum oocysts, with various ratios of genotype I to genotype II, were isolated and subjected to PCR-restriction fragment length polymorphism analysis. Amplification of both genotypes in these samples ranged from 31 to 74% and yielded no information ...
In the present study, a robust TaqMan real-time PCR amplifying the F57 and the ISMav2 sequences of Mycobacterium avium subsp. paratuberculosis from bovine fecal samples was developed and validated. The validation was based on the recommendations of International Organization for Standardization protocols for PCR and real-time PCR methods. For specificity testing, 205 bacterial strains were selected, including 105 M. avium subsp. paratuberculosis strains of bovine, ovine, and human origin and 100 non-M. avium subsp. paratuberculosis strains. Diagnostic quality assurance was obtained by use of an internal amplification control. By investigating six TaqMan reagents from different suppliers, the 100% detection probability was assessed to be 0.1 picogram M. avium subsp. paratuberculosis DNA per PCR. The amplification efficiency was 98.2% for the single-copy gene F57 and 97.8% for the three-copy insertion sequence ISMav2. The analytical method was not limited due to instrument specificity. The triplex real
TY - JOUR. T1 - Detection of equine arteritis virus by real-time TaqMan® reverse transcription-PCR assay. AU - Balasuriya, Udeni B R. AU - Leutenegger, Christian M.. AU - Topol, J. B.. AU - McCollum, William H.. AU - Timoney, Peter J.. AU - Maclachlan, Nigel J. PY - 2002. Y1 - 2002. N2 - A one-tube real-time TaqMan® reverse transcription-polymerase chain reaction (RT-PCR) assay was developed for the detection of equine arteritis virus (EAV). The test was validated using the seminal plasma and nasal secretions of infected horses that were proven to contain EAV by traditional virus isolation in rabbit kidney thirteen (RK-13) cells, as well as a variety of cell culture-propagated European and North American strains of EAV. The primers and a fluorogenic TaqMan® probe were designed to amplify and detect a highly conserved region of open reading frame 7 (ORF7) of EAV. The real-time TaqMan® PCR assay detected EAV RNA in all samples that were confirmed to contain infectious EAV by virus isolation. ...
Microsatellite markers derived from simple sequence repeats have been useful in studying a number of human pathogens, including the human malaria parasite Plasmodium falciparum. Genetic markers for P. vivax would likewise help elucidate the genetics and population characteristics of this other important human malaria parasite. We have identified a locus in a P. vivax telomeric clone that contains simple sequence repeats. Primers were designed to amplify this region using a two-step semi-nested polymerase chain reaction protocol. The primers did not amplify template obtained from non-infected individuals, nor DNA from primates infected with the other human malaria parasites (P. ovale, P. malariae, or P. falciparum). The marker was polymorphic in P. vivax-infected field isolates obtained from Papua New Guinea, Indonesia and Guyana. This microsatellite marker may be useful in genetic and epidemiologic studies of P. vivax malaria.
Despite the rapid development of pharmacogenetic testing and the frequencies of medication related emergencies increasing, pharmacogentic testing is not yet implemented extensively in clinical practice. Several studies document that polymorphic Cytochrome P450 isoenzymes CYP2C9, CYP2C19 and CYP2D6 are responsible for the metabolism of many clinically important drugs and xenobiotics. These polymorphisms contribute to inter-individual and interethnic variation in drug metabolism which may lead to differences in drug response, suggesting that common dose regimen will not always equivocate to efficacious dosing. The CYP2D6*2, CYP2D6*4 and CYP2D6*10 variants were studied in four Indian populations (Gujarati, Punjabi, Bengali and Koya tribe). Genotypes at individual alleles were identified using Polymerase Chain Reaction Restriction Fragment Length Polymorphism (PCR-RFLP) method. Differences in frequencies of CYP2D6 genotypes/ alleles were compared at regional and global level. CYP2D6*2 variant ...
Polymorphisms in paraoxonase 1 (PON1) coding for PON1 enzyme have been studied as genetic markers of coronary artery disease (CAD). PON1 Q192R and PON1 L55M polymorphisms have been analyzed extensively, but data on association and role of these polymorphisms in the etiology of CAD are conflicting. In this study, we tested the genetic association between PON1 Q192R and PON1 L55M polymorphisms and CAD among north Indians. MATERIALS AND METHODS: Two hundred eighty-five angiographically proven patients with coronary artery disease and 200 sex-matched and ethnically matched controls were genotyped for 2 PON1 polymorphisms by the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technique. Genotype/ allele frequencies were compared in patients and controls using the chi-square test. RESULTS: At PON1-192 locus, there were significant differences between patients and controls (P, 0.05), leading to significant odds ratios for RR genotype (OR= 1.92, CI: 1.19-3.10) and *R allele ...
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 (DNA). DNA profiling is today possible with even very small quantities of blood: this is commonly ...
Main articles: Taq Polymerase and History of polymerase chain reaction. In 1983, Mullis was working for Cetus Corporation as a ... Invention of polymerase chain reaction. Awards. William Allan Award (1990). Robert Koch Prize (1992). Nobel Prize in Chemistry ... K. Mullis, 1990, The unusual origin of the polymerase chain reaction. Scientific American, April 56-65. ... The Polymerase Chain Reaction, 1994, co-edited Francious Ferre and Richard A. Gibbs (Basel: Birkhauser) .mw-parser-output cite. ...
Polymerase Chain Reaction: An Alternative to Cloning *^ Brown TA (2002). "Section 2, Chapter 6: 6.1. The Methodology for DNA ... DNA can also be amplified using a procedure called the polymerase chain reaction (PCR).[93] By using specific short sequences ... Kary Banks Mullis developed the polymerase chain reaction, providing a quick way to isolate and amplify a specific section of ... Each strand of DNA is a chain of nucleotides, matching each other in the center to form what look like rungs on a twisted ...
A polymerase chain reaction (PCR) test has been developed for the detection of Babesia from the peripheral blood.[12] PCR may ... "Detection of Babesia microti by polymerase chain reaction". J. Clin. Microbiol. 30 (8): 2097-103. PMC 265450. PMID 1500517.. ...
Weier, HU; Gray, JW (Jul-Aug 1988). "A programmable system to perform the polymerase chain reaction". DNA (Mary Ann Liebert, ... is a laboratory apparatus most commonly used to amplify segments of DNA via the polymerase chain reaction (PCR).[1] Thermal ... Later, the PCR process was adapted to the use of thermostable DNA polymerase from Thermus aquaticus, which greatly simplified ... The earliest thermal cyclers were designed for use with the Klenow fragment of DNA polymerase I. Since this enzyme is destroyed ...
Polymerase chain reactionEdit. Main article: Polymerase chain reaction. Polymerase chain reaction (PCR) is an extremely ... "Polymerase Chain Reaction (PCR) Fact Sheet". National Human Genome Research Institute (NHGRI). Retrieved 31 December 2016.. ... "Polymerase Chain Reaction (PCR)". www.ncbi.nlm.nih.gov. Retrieved 31 December 2016.. ... In this technique, DNA coding for a protein of interest is cloned using polymerase chain reaction (PCR), and/or restriction ...
Another method of screening is with polymerase chain reaction (PCR). Some libraries are stored as pools of clones and screening ...
Polymerase chain reaction(PCR).[44] Additionally the development of recombinant DNA technology through the use of bacteria has ... From this microbe scientists isolated the enzyme Taq polymerase, which is now used in the powerful experimental technique, ... Fröls S, White MF, Schleper C (2009). "Reactions to UV damage in the model archaeon Sulfolobus solfataricus". Biochemical ... Terpe, Kay (1 November 2013). "Overview of thermostable DNA polymerases for classical PCR applications: from molecular and ...
Polymerase chain reaction is done by placing a mixture of the desired DNA, DNA polymerase, primers, and nucleotide bases into a ... Molecular biology consists of different techniques including Polymerase chain reaction, Gel electrophoresis, and macromolecule ... Proteins are chains of amino acids that function to contract skeletal muscle, function catalysts, transport molecules, and ... Proteins can facilitate biochemical processes, by lowering the activation energy of a reaction. Hemoglobins are also proteins, ...
for the development of the polymerase chain reaction. 1992 格哈德·埃特爾. 德國 for the contributions to the new development of the ... for the discovery of Immunoglobulin E and mechanisms of IgE-mediated allergic reactions. ...
... polymerase chain reaction) helps DNA polymerase to create new strands of DNA equivalent to template given.[17] ... "Polymerase Chain Reaction (PCR)". National Center for Biotechnology Information. Retrieved 2016-06-10.. ... but patients treated with paromomycin had a higher rate of adverse skin reactions.[19] ...
Alternatively, diagnosis and strain identification can be made using polymerase chain reaction (PCR).[15] ...
Tests that use polymerase chain reaction (PCR, aka nucleic acid amplification) to identify genes unique to N. gonorrhoeae are ... Traditionally, gonorrhea was diagnosed with Gram stain and culture; however, newer polymerase chain reaction (PCR)-based ... It is also possible for an individual to have an allergic reaction to the bacteria, in which case any appearing symptoms will ...
Wilks (1989). "Two putative protein-tyrosine kinases identified by application of the polymerase chain reaction". PNAS. 86 (5 ...
... for his invention of the polymerase chain reaction (PCR) method; for contributions to the developments of methods within DNA- ... "for their work on chirally catalysed hydrogenation reactions; for his work on chirally catalysed oxidation reactions". ... "for their theories, developed independently, concerning the course of chemical reactions". *^ "The Nobel Prize in Chemistry ... "for their studies of extremely fast chemical reactions, effected by disturbing the equilibrium by means of very short pulses of ...
380-nL reaction chambers, and capillary electrophoresis channels for reverse transcription polymerase chain reaction (RT-PCR) ... The polymerase chain reaction (PCR) is a fundamental molecular biology technique that enables the selective amplification of ... Toriello, Nicholas M.; Liu, Chung N.; Mathies, Richard A. (2006). "Multichannel Reverse Transcription-Polymerase Chain Reaction ... the first continuous-flow polymerase chain reaction chip was developed In 1999, the first demonstration of heterogeneous ...
Fouchier used a broad-spectrum "pan-coronavirus" real-time polymerase chain reaction (RT-PCR) method to test for distinguishing ... "Detection of a novel human coronavirus by real-time reverse-transcription polymerase chain reaction". Euro Surveillance. 17 (39 ... as well as for RNA-dependent RNA polymerase (RdRp), a gene conserved in all coronaviruses known to infect humans. While the ...
Effects of Heparin on Polymerase Chain Reaction for Blood White Cells. J. Clin. Lab. Anal. 1999, 13 (3): 133-140. PMID 10323479 ... Outbreak of adverse reactions associated with contaminated heparin. N Engl J Med. 2008, 359 (25): 2674-84. PMC 3810025. PMID ...
H7 from filth flies by polymerase chain reaction". Medical and Entomology. 18 (3): 241-246. doi:10.1111/j.0269-283X.2004.00502. ...
Several different polymerase chain reaction (PCR) tests are available for the detection of Leishmania DNA.[3] With this assay, ... RNA polymerase II transcribes long polycistronic messages in the absence of defined RNA pol II promoters, and Leishmania has ...
Blood samples can be sent away for polymerase chain reaction (PCR) testing to confirm infection. Other diseases that might ... "Dectection of Persistent Cytauxzoon Felis Infection by Polymerase Chain Reaction in Three Asymptomatic Domestic Cats". Journal ...
Polymerase chain reaction (PCR) tests where available, mostly in industrialized countries, have been increasingly used; PCR can ... catalase for which all clinically relevant Neisseria show a positive reaction, and the carbohydrates maltose, sucrose, and ...
In children, polymerase chain reaction sequencing of urine can detect fragments of the infectious agent. The procedure differs ...
placebo . plant nutrition . plasmolysis . pollination . polygene . polymer . polymerase chain reaction . polyploid . population ... an endergonic reaction (also called a nonspontaneous reaction or an unfavorable reaction) is a chemical reaction in which the ... chemical kinetics The study and discussion of chemical reactions with respect to reaction rates, effect of various variables, ... flagellum . flavin adenine dinucleotide . food chain The chain of eating and getting nutrition which starts from a small ...
The claim itself has two simple and conventional steps: first amplifying (by polymerase chain reaction, PCR) and then detecting ...
"Diagnosis of primary human herpesvirus 6 and 7 infections in febrile infants by polymerase chain reaction". Archives of Disease ... "Diagnosis of primary human herpesvirus 6 and 7 infections in febrile infants by polymerase chain reaction". Archives of Disease ... of a PCR system for porcine cytomegalovirus detection and determination of the putative partial sequence of its DNA polymerase ...
... ... Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP) is a method with the addition of the enzyme after ... Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP) merupakan metode PCR dengan penambahan enzim ... Objective: To determine the diagnostic value of Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP) ...
Sexing of Sheep Embryos Produced In vitro by Polymerase Chain Reaction and Sex-specific Polymorphism2003 January;16(5) ... Identification of Pork Contamination in Meatballs of Indonesia Local Market Using Polymerase Chain Reaction-Restriction ... Characterization of MHC DRB3.2 Alleles of Crossbred Cattle by Polymerase Chain Reaction-Restriction Fragment Length ... Characterization of MHC DRB3.2 Alleles of Crossbred Cattle by Polymerase Chain Reaction-Restriction Fragment Length ...
End of Molecular Pathology , Polymerase Chain Reaction , Taq polymerase. This information is intended for physicians and ... Additionally, Taq often adds an adenine to the end of its polymerase reaction (sometimes used to facilitate TA cloning) ... Initially the Klenow fragment from the E. coli DNA polymerase I was used in PCR reactions ... The polymerase, known as the Taq polymerase, has many attributes that are well suited for PCR: ...
... ( 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 ...
Scientific American is the essential guide to the most awe-inspiring advances in science and technology, explaining how they change our understanding of the world and shape our lives.
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. ...
Polymerase Chain Reaction Detection of Herpes Simplex Virus in Cerebrospinal Fluid R. H. Boerman, A. C. B. Peters, E. P. J. ... Polymerase Chain Reaction Diagnosis of Varicella Zoster Virus Ravi Mahalingam, Randall Cohrs, Aud N. Dueland, Donald H. Gilden ... Polymerase Chain Reaction for Hepatitis Delta Virus RNA Identification and Characterization Anna Linda Zignego, Paul Deny, ... Antigen Capture/Polymerase Chain Reaction for the Detection of Hepatitis A Virus in Human Clinical Materials ...
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 ...
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 ...
Specific detection of monkeypox virus by polymerase chain reaction.. Neubauer H1, Reischl U, Ropp S, Esposito JJ, Wolf H, Meyer ...
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 ...
... 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 ...
An optical instrument monitors PCR replication of DNA in a reaction apparatus having a temperature cycled block with vials of ... reaction ingredients including dye that fluoresces in presence of double-stranded DNA. A beam splitter passes an excitation ... A filter module for an optical instrument that monitors polymerase chain reaction replication of DNA in a reaction apparatus ... 1. An optical instrument for monitoring polymerase chain reaction replication of DNA in a reaction apparatus that includes a ...
This study aimed to establish an accurate and sensitive polymerase chain reaction [‎PCR]‎ technique for the diagnosis of active ... Diagnosis of human brucellosis in Egypt by polymerase chain reaction. EMHJ - Eastern Mediterranean Health Journal, 15 (‎5)‎, ...
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 ...
  • Specific detection of monkeypox virus by polymerase chain reaction. (nih.gov)
  • Reactions were carried out on the ABI PRISM® 7700 Sequence Detection Systems. (sigmaaldrich.com)
  • Detection of Burkholderia pseudomallei in blood samples using polymerase chain reaction. (nih.gov)
  • Detection of bifidobacteria can be achieved using DNA extracted from human faeces as template in PCR reactions. (scribd.com)
  • To obtain standardized quantitative results, external controls consisting of a DNA template for each target gene were used to generate linear standard curves over a 6 to 8 log range with detection of as few as 10 copies of amplicon per reaction. (bioone.org)
  • Detection of specific polymerase chain reaction product by utilizing the 5'----3' exonuclease activity of Thermus aquaticus DNA polymerase. (pnas.org)
  • Nucleic acid hybridization techniques, exploited by colony hybridizations or polymerase chain reaction (PCR), apply a single detection method to a diversity of organisms. (cdc.gov)
  • Detection of Mycoplasma pneumoniae in clinical samples from pediatric patients by polymerase chain reaction. (asm.org)
  • Polymerase chain reaction (PCR) for the detection of food-borne pathogens. (ihs.com)
  • Reverse-transcriptase-polymerase chain reaction (RT-PCR) has been applied successfully for detection of canine distemper virus (Agnihotri et al. (thefreelibrary.com)
  • After dissection in search of flagellates in digestive tubes and identification of the species, female sandflies were submitted to the Multiplex Polymerase Chain Reaction (multiplex PCR) for detection of the fragment of the kDNA of Leishmania (Viannia) and the fragment from the IVS6 cacophony gene region of the phlebotomine insects. (scielo.br)
  • The Application Analysis of Multiplex Real-Time Polymerase Chain Reaction Assays for Detection of Pathogenic Bacterium in Peritoneal Dialysis-Associated Peritonitis. (bioportfolio.com)
  • To estimate the clinical value of bacterial detection in peritoneal dialysis-associated peritonitis (PDAP) by multiplex real-time polymerase chain reaction (RT-PCR). (bioportfolio.com)
  • Detection of cytomegalovirus (CMV) DNA by real-time polymerase chain reaction (rt-PCR) in dried blood spots (DBS) collected for newborn screening has been assessed for retrospective diagnosis of conge. (bioportfolio.com)
  • Detection of the Lyme disease bacterium, Borrelia burgdorferi, by using the polymerase chain reaction and a nonradioisotopic gene probe. (prohealth.com)
  • Detection of viable but nonculturable Vibrio parahaemolyticus induced by prolonged cold-starvation using propidium monoazide real-time polymerase chain reaction. (bioportfolio.com)
  • The aim of this study was to develop a nested polymerase chain reaction (nested PCR) for the rapid detection of transmissible gastroenteritis virus (TGEV) of pigs. (eurekamag.com)
  • The polymerase chain reaction (PCR) was evaluated for the detection of leptospires in clinical samples from patients with acute leptospiral infection. (microbiologyresearch.org)
  • We detected folate receptor (FR)-positive circulating tumor cells (FR + -CTCs) by a novel ligand-targeted polymerase chain reaction (LT-PCR) detection technique. (jcancer.org)
  • 263 Detection of gonadotropin-releasing hormone in the rat ovary : studies using the reverse transcription-polymerase chain reaction. (go.jp)
  • We wished to develop an accurate quantitative protocol based on a droplet digital polymerase chain reaction (ddPCR) involving eight individual TaqManreactions to detect simultaneously, without selective enrichment, Listeria spp. (frontiersin.org)
  • Analysis of major BCR-ABL1 mRNA by digital polymerase chain reaction is useful for prediction of international scale. (bioportfolio.com)
  • 9. A method in accordance with claim 1 wherein said first subset of PCR reagents consists of all PCR reagents except a nucleic acid polymerase. (google.es)
  • In Situ Polymerase Chain Reaction ( In situ PCR) is a powerful method that detects minute quantities of rare or single-copy number nucleic acid sequences in frozen or paraffin-embedded cells or tissue sections for the localization of those sequences within the cells. (biogenex.com)
  • With traditional PCR, the amount of product can only be worked out at the end of the reaction (the plateau phase), typically by agarose gel electrophoresis. (abpischools.org.uk)
  • Mycobacterium tuberculosis isolates from previously treated patients (n = 88) from all regions of Syrian Arab Republic were characterized in terms of antibiotic sensitivity and genotyping using double-repetitive-element polymerase chain reaction (DRE-PCR) method for the proximity of the repetitive DNA elements IS6110 (a mobile genetic element) and PGRS. (who.int)
  • In the present study, a genotyping method based on polymerase chain reaction (PCR) for the repeat regions of four genes (sdrG, sdrF, aap and sesE) that encode for bacterial surface proteins was developed and applied to a sample of well-characterised neonatal blood isolates of S. epidermidis (n = 49). (diva-portal.org)
  • Polymerase chain reaction analysis of two biopsy tissues using the Cobas TaqMan revealed a positive result for Mycobacterium avium and a negative result for Mycobacterium tuberculosis . (hindawi.com)
  • However, polymerase chain reaction analysis of a cultured colony of acid-fast bacteria from biopsy tissue using the Cobas TaqMan and an alternative polymerase chain reaction analysis of biopsy tissue yielded discordant results. (hindawi.com)
  • TaqMan polymerase chain reaction was developed to quantify the number of Bifidobacterium . (ajol.info)
  • Two polymerase chain reaction (PCR) protocols showed low sensitivity (36% and 53% for TB AMPLICOR and MPB64 nested PCR, respectively), when compared with classic microbiological methods (73% and 54% for Ziehl-Neelsen staining and culture, respectively), in the diagnosis of tuberculous meningitis in 91 patients in southeastern Brazil. (scielo.br)
  • The diagnosis of tuberculous meningitis using the polymerase chain reaction. (nus.edu.sg)
  • MilliporeSigma's molecular biologists work to develop the newest, best performing polymerases for customer use, optimizing conditions, buffer compositions, and cycling parameters to save you valuable time and resources. (emdmillipore.com)
  • If the polymerase used was heat-susceptible, it would denature under the high temperatures of the denaturation step. (wikipedia.org)
  • Recently, simple and rapid assays for quantifying mRNA expression by real-time reverse transcription-polymerase chain reaction (RT-PCR) have been used for analysis of cytokine profiles in humans and other mammalian species. (bioone.org)
  • Reverse transcription polymerase chain reaction, abbreviated RT-PCR, detects mRNA, pre-mRNA and noncoding RNAs. (golden.com)
  • Major BCR-ABL1 mRNA in patients with chronic myeloid leukemia (CML) has generally been analysed by real-time polymerase chain reaction (PCR). (bioportfolio.com)
  • Furthermore, automation has been facilitated, as reaction buffers and additional reagents can be rapidly changed without centrifugation or precipitation steps. (springer.com)
  • The purpose of this study is to demonstrate the efficacy of active study vaccine in the prevention of reverse transcriptase polymerase chain reaction (RT-PCR) confirmed respiratory syncytial virus (RSV)-mediated lower respiratory tract disease (LRTD), when compared to placebo. (centerwatch.com)
  • BACKGROUND Real-time reverse transcription polymerase chain reaction (RT-PCR) is routinely used to detect viral infections. (bireme.br)
  • Comparative Evaluation of Immunochromatographic and Reverse Transcriptase Polymerase Chain Reaction based tests for Diagnosis of Canine Distemper. (thefreelibrary.com)
  • An analysis of the transcript level of the 10 members of the family has been performed using the technique of real-time quantitative reverse transcriptase-polymerase chain reaction. (plantphysiol.org)
  • Reverse transcriptase polymerase chain reaction (RT-PCR) can detect CTCs with high sensitivity [ 8 ]. (jcancer.org)
  • Introduction The polymerase chain reaction (PCR) is a method that uses test tubes in biological laboratories for producing large amount of identical copies of a specific gene from small amount of complex molecules. (psu.edu)
  • Polymerase chain reaction ( PCR ) is a method widely used in molecular biology to make many copies of a specific DNA segment. (wikipedia.org)
  • A mainstay of virtually every molecular biology lab, polymerase chain reaction (PCR) is an easy and affordable method for amplifying specific fragments of DNA by several orders of magnitude. (sigmaaldrich.com)
  • A PCR or polymerase chain reaction is a laboratory procedure in which millions of copies of a specific piece of DNA are made. (myvmc.com)
  • The contamination of Taq polymerase by bacterial DNA is now well established in the published press. (bmj.com)
  • In order to try to assess the extent of inhibition that occurs in a reaction, a control can be performed by adding a known amount of a template to the investigated reaction mixture (based on the sample under analysis). (wikipedia.org)
  • DNA sample preparation, reaction mixture assemblage and the PCR process, in addition to the subsequent reaction product analysis, should be performed in separate areas. (stanford.edu)
  • A Laminar Flow Cabinet equipped with a UV lamp is recommended for preparing the reaction mixture. (stanford.edu)
  • The use of dedicated vessels and positive displacement pipettes or tips with aerosol filters for both DNA sample and reaction mixture preparation, is strongly recommended. (stanford.edu)
  • Usually the amount of template DNA is in the range of 0.01-1 ng for plasmid or phage DNA and 0.1-1 µg for genomic DNA , for a total reaction mixture of 50 µl. (stanford.edu)
  • DNA-based procedures are becoming increasingly common within the analytical laboratory where the polymerase chain reaction (PCR) has become an indispensable technique. (rsc.org)
  • To standardise and apply nested Polymerase Chain Reaction (n PCR) as a rapid and reliable laboratory diagnostic test to detect Toxoplasma gondii (T.gondii) in aqueous humor (AH) and peripheral blood samples of clinically suspected toxoplasma chorioretinitis patients. (arvojournals.org)
  • To order a free set of ABPI schools posters on the following topics: biotechnology, cloning, genetic engineering, unravelling the genome, polymerase chain reaction and stem cells, please fill in our order form . (abpischools.org.uk)
  • MS2-like particles were produced through the synthesis of MS2 RNA genome by T7 RNA polymerase. (bireme.br)
  • The polymerase chain reaction, now widely used in research laboratories and doctor's offices, relies on the ability of DNA-copying enzymes to remain stable at high temperatures. (accessexcellence.org)
  • When any cell divides, enzymes called polymerases make a copy of all the DNA in each chromosome. (accessexcellence.org)
  • KOD polymerase yields more product in fewer cycles compared to other PCR enzymes. (emdmillipore.com)
  • In the case of RT-PCR, these are labelled with a fluorescent reporter dye at the 5' end and a quencher dye at the 3' end and only produce fluorescence when DNA polymerase moves along the template and cleaves (splits) the probe. (abpischools.org.uk)
  • As the DNA polymerase moves along the template, the probe is cleaved (broken) between the reporter and quencher dye. (abpischools.org.uk)
  • Recently, the application of the polymerase chain reaction (peR) technique developed by K. D. Mullis and detailed in the study by Saiki et al. (springer.com)
  • PCR (Polymerase chain reaction) was developed by Kary Mullis in the mid-1980s. (slideserve.com)
  • The KCl concentration (30 to 90 mM) implied no significant effect to the yields of PCR products when higher Taq polymerase (1.0 to 2.5 units/40 μl total reaction) was included. (iwaponline.com)
  • Lorenz, T. C. Polymerase Chain Reaction: Basic Protocol Plus Troubleshooting and Optimization Strategies. (jove.com)
  • 2014). This study is an attempt to compare polymerase chain reaction based molecular tests with commercially available IC based test for diagnosis of Canine distemper by testing rectal swabs and serum samples respectively from dogs suffering from gastroenteritis and suspected for Canine distemper. (thefreelibrary.com)
  • Between May and October 2015 we analysed the serum samples of 140 patients with a suspicion of dengue, using ELISA and multiplex polymerase chain reaction. (who.int)
  • A control reaction, omiting template DNA, should always be performed, to confirm the absence of contamination. (stanford.edu)
  • The most sensitive quantification methods are done by the real-time polymerase chain reaction , where the amount of DNA is measured after each cycle of PCR by use of fluorescent markers. (bionity.com)
  • Due to limitations of conventional diagnostic methods, polymerase ch. (bioportfolio.com)
  • We aimed to evaluate the impact of PCVs on invasive pneumococcal disease (IPD) using polymerase chain reaction (PCR)-based techniques and compare with results obtained from culture-based methods. (biomedcentral.com)