PCR protocols : a guide to methods and applications


PCR protocols : a guide to methods and applications

edited by Michael A. Innis ... [et al.]

Academic Press, c1990

  • : hbk
  • : pbk

大学図書館所蔵 件 / 111



Includes bibliographies and index



: hbk ISBN 9780123721808


The correct procedures you need for frustration-free PCR methods and applications are contained in this complete, step-by-step, clearly written, inexpensive manual.


[Contents in a condensed version] Basic Methodology Amplification of genomic DNA targets Optimization of PCR reaction and design of primers Nonradioactive detection of PCR products mRNA quantitation by PCR Cloning PCR products Contamination and product carry-over PCR from paraffin sections Amplifying ancient DNA Research Applications Direct DNA sequencing of PCR products DNA footprinting Detection of rare mRNAs Site-directed mutagenesis Screening DNA libraries Amplification of flanking DNA sequences Genetics and Evoltion Tissue typing and disease susceptibility Haplotype analysis of single cells Population biology Multiplex DNA amplification Diagnostics and Forensics HIV detection HTLV I and II detection Screening for novel viruses Detection of oncogenic mutations Diagnosis of retinoblastoma Individual identification Instruments and Supplies Organizing a laboratory for PCR work Low-cost air-driven cycling oven Basic equipment and supplies for the PCR lab

: pbk ISBN 9780123721815


The broad utility of the polymerase chain reaction (PCR) method is now within the reach of every researcher. Designed for use at the laboratory bench, this is the most comprehensive manual on PCR available today. Over 50 chapters provide precise instructions on procedures, with advice on primer design. All of the techniques described, from amplification and direct sequencing of genomic DNA through cDNA cloning and quantitation of mRNA, are tested, current, and supplemented with helpful notes and illustrations. You'll also learn how to: optimize novel applications; avoid many cumbersome molecular biological techniques; and set up the laboratory to avoid contamination. This first-rate guide will help you: avoid contamination - with specific instructions on setting up your lab; avoid cumbersome molecular biological techniques; and, discover new applications.


  • 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.

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