Topic
Polymerase chain reaction
About: Polymerase chain reaction is a research topic. Over the lifetime, 18408 publications have been published within this topic receiving 768719 citations. The topic is also known as: PCR.
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TL;DR: A new DNA polymorphism assay based on the amplification of random DNA segments with single primers of arbitrary nucleotide sequence is described, suggesting that these polymorphisms be called RAPD markers, after Random Amplified Polymorphic DNA.
Abstract: Molecular genetic maps are commonly constructed by analyzing the segregation of restriction fragment length polymorphisms (RFLPs) among the progeny of a sexual cross. Here we describe a new DNA polymorphism assay based on the amplification of random DNA segments with single primers of arbitrary nucleotide sequence. These polymorphisms, simply detected as DNA segments which amplify from one parent but not the other, are inherited in a Mendelian fashion and can be used to construct genetic maps in a variety of species. We suggest that these polymorphisms be called RAPD markers, after Random Amplified Polymorphic DNA.
13,764 citations
Book•
01 Jan 1990
TL;DR: Basic Methodology: M.A. Innis and D.F. Frohman, RACE: Rapid Amplification of cDNA Ends, and RNA Processing: Apo-B.R. Kwok, Procedure to Minimuze PCR-Product Carry-Over.
Abstract: Basic Methodology: M.A. Innis and D.H. Gelfand, Optimization of PCRs. R.K. Saiki, Amplification of Genomic DNA. E.S. Kawasaki, Amplification of RNA. M.A. Frohman, RACE: Rapid Amplification of cDNA Ends. T. Compton, Degenerate Primers for DNA Amplification. C.C. Lee and C.T. Caskey, cDNA Cloning Using Degenerate Primers. M.A. Innis, PCR with 7-Deaza-2~b7-Deoxyguanosine Triphosphate. G. Gilliland, S. Perrin, and H.F. Bunn, Competitive PCR for Quantitation of mRNA. A.M. Wang and D.F. Mark, Quantitative PCR. P.C. McCabe, Production of Single-Stranded DNA by Asymmetric PCR. S.J. Scharf, Cloning with PCR. U. Landegren, R. Kaiser, and L. Hood, Oligonucleotide Ligation Assay. C. Levenson and C.-A. Chang, Nonisotopically Labeled Probes and Primers. Y-M.D. Lo, W.Z. Mehal, and K.A. Fleming, Incorporation of Biotinylated dUTP. R. Helmuth, Nonisotopic Detection of PCR Products. D.H. Gelfand and T.J. White, Thermostable DNA Polymerases. S. Kwok, Procedure to Minimuze PCR-Product Carry-Over. E.S. Kawasaki, Sample Preparation from Blood, Cells, and Other Fluids. D.K. Wright and M.M. Manos, Sample Preparation from Paraffin-Embedded Tissues. S. P~ada~adabo, Amplifying Ancient DNA. Research Applications. M.J. Holland and M.A. Innis, In Vitro Transcription of PCR Templates. R. Higuchi, Recombinant PCR. B. Krummel, DNase I Footprinting. M.A.D. Brow, Sequencing with Taq DNA Polymerase. S.S. Sommer, G. Sarkar, D.D. Koeberl, C.D.K. Bottema, J.-M. Buerstedde, D.B. Schowalter, and J.D. Cassady, Direct Sequencing with the Aid of Phage Promoters. V.C. Sheffield, D.R. Cox, and R.M. Myers, Identifying DNA Polymorphisms by Denaturing Gradient Gel Electrophoresis. H. Ochman, M.M. Medhora, D. Garza, and D.L. Hartl, Amplification of Flanking Sequences by Inverse PCR. M.A. Frohman and G.R. Martin, Detection of Homologous Recombinants. L.M. Powell, RNA Processing: Apo-B. T.R. Gingeras, G.R. Davis, K.M. Whitfield, H.L. Chappelle, L.J. DiMichele, and D.Y. Kwoh, A Transcription-Based Amplification System. K.D. Friedman, N.L. Rosen, P.J. Newman, and R.R. Montgomery, Screening of ~glgt11 Libraries. Genetics and Evolution. H.A. Erlich and T.L. Bugawan, HLA DNA Typing. J.S. Chamberlain, R.A. Gibbs, J.E. Ranier, and C.T. Caskey, Multiplex PCR for the Diagnosis of Duchenne Muscular Dystrophy. S.B. Lee and J.W. Taylor, Isolation of DNA from Fungal Mtcelia and Single Spores. S.C. Kogan and J. Gitschier, Genetic Prediction of Hemophilia A. U. Gyllensten, Haplotype Analysis from Single Sperm or Diploid Cells. M.L. Sogin, Amplification of Ribosomal RNA Genes for Molecular Evolution Studies. T.J. White, T. Bruns, S. Lee, and J. Taylor, Amplification and Direct Sequencing of Fungal Ribosomal RNA Genes for Phylogenetics. Diagnostics and Forensics. G.D. Ehrlich, S. Greenberg, and M.A. Abbott, Detection of Human T-Cell Lymphoma/Leukemia Viruses. D.E. Kellogg and S. Kwok, Detection of Human Immunodeficiency Virus. I. Baginski, A. Ferrie, R. Watson, and D. Mack, Detection of Hepatitis B Virus. Y. Ting and M.M. Manos, Detection and Typing of Genital Human Papillomaviruses. D. Shibata, Detection of Human Cytomegalovirus. H.A. Rotbart, PCR Amplification of Enteroviruses. D. Mack, O.-S. Kwon, and F. Faloona, Novel Viruses. J. Lyons, Analysis of ras Gene Point Mutations by PCR and Olgonucleotide Hybridization. M. Crescenzi, B-Cell Lymphoma: t(14 18) Chromosome Rearrangement. R.M. Atlas and A.K. Bej, Detecting Bacterial Pathogens in Environmental Water Samples by Using PCR and Gene Probes. S.-H. Park, PCR in the Diagnosis of Retinoblastoma. C. Orrego and M.C. King, Determination of Familial Relationships. Instrumentation and Supplies: R. Watson, PCR in a Teacup A Simple and Inexpensive Method for Thermocycling PCRs. P. Denton and H. Reisner, A Low-Cost Air-Driven Cycling Oven. N.C.P. Cross, N.S. Foulkes, D. Chappel, J. McDonnell, and L. Luzzatto, Modification of a Histokinette for Use as an Automated PCR Machine. C. Orrego, Organizing a Laboratory for PCR Work. R. Madej and S. Scharf, Basic Equipment and Supplies. Index.
13,139 citations
TL;DR: Analysis of the genomic DNA from a bacterial biofilm grown under aerobic conditions suggests that sulfate-reducing bacteria, despite their anaerobicity, were present in this environment.
Abstract: We describe a new molecular approach to analyzing the genetic diversity of complex microbial populations. This technique is based on the separation of polymerase chain reaction-amplified fragments of genes coding for 16S rRNA, all the same length, by denaturing gradient gel electrophoresis (DGGE). DGGE analysis of different microbial communities demonstrated the presence of up to 10 distinguishable bands in the separation pattern, which were most likely derived from as many different species constituting these populations, and thereby generated a DGGE profile of the populations. We showed that it is possible to identify constituents which represent only 1% of the total population. With an oligonucleotide probe specific for the V3 region of 16S rRNA of sulfate-reducing bacteria, particular DNA fragments from some of the microbial populations could be identified by hybridization analysis. Analysis of the genomic DNA from a bacterial biofilm grown under aerobic conditions suggests that sulfate-reducing bacteria, despite their anaerobicity, were present in this environment. The results we obtained demonstrate that this technique will contribute to our understanding of the genetic diversity of uncharacterized microbial populations.
11,380 citations
TL;DR: A set of oligonucleotide primers capable of initiating enzymatic amplification (polymerase chain reaction) on a phylogenetically and taxonomically wide range of bacteria is described in this paper.
Abstract: A set of oligonucleotide primers capable of initiating enzymatic amplification (polymerase chain reaction) on a phylogenetically and taxonomically wide range of bacteria is described along with methods for their use and examples. One pair of primers is capable of amplifying nearly full-length 16S ribosomal DNA (rDNA) from many bacterial genera; the additional primers are useful for various exceptional sequences. Methods for purification of amplified material, direct sequencing, cloning, sequencing, and transcription are outlined. An obligate intracellular parasite of bovine erythrocytes, Anaplasma marginale, is used as an example; its 16S rDNA was amplified, cloned, sequenced, and phylogenetically placed. Anaplasmas are related to the genera Rickettsia and Ehrlichia. In addition, 16S rDNAs from several species were readily amplified from material found in lyophilized ampoules from the American Type Culture Collection. By use of this method, the phylogenetic study of extremely fastidious or highly pathogenic bacterial species can be carried out without the need to culture them. In theory, any gene segment for which polymerase chain reaction primer design is possible can be derived from a readily obtainable lyophilized bacterial culture.
10,245 citations
TL;DR: Six primers for the amplification of three non-coding regions of chloroplast DNA via the polymerase chain reaction (PCR) have been designed and worked for most species tested, which means that they may be used to study the population biology and evolution of plants.
Abstract: Six primers for the amplification of three non-coding regions of chloroplast DNA via the polymerase chain reaction (PCR) have been designed. In order to find out whether these primers were universal, we used them in an attempt to amplify DNA from various plant species. The primers worked for most species tested including algae, bryophytes, pteridophytes, gymnosperms and angiosperms. The fact that they amplify chloroplast DNA non-coding regions over a wide taxonomic range means that these primers may be used to study the population biology (in supplying markers) and evolution (inter- and probably intraspecific phylogenies) of plants.
5,212 citations