This manual introduces the complex world of PCR by beginning at an accessible level and then moving to more advanced levels of application. First, the practical requirements for performing PCR and other amplification techniques in the lab are introduced and then the basic aspects of the technique are explained by exploring important issues such as sample preparation, primer design, efficiency, detection of products, and quantitation. Protocols for a range of PCR and amplification techniques are presented for cloning, sequencing, mutagenesis, library construction and screening, exon trapping, differential display, and expression, and these include RT-PCR, RNA PCR, LCR, multiplex PCR, panhandle PCR, capture PCR, expression PCR, 3' and 5' RACE, in situ PCR, and litigation-mediated PCR. Each protocol is augmented by analysis and troubleshooting sections and complete references.

Table of Contents:
Part 1 Introduction to PCR: setting up a PCR laboratory, C.W. Dieffenbach et al; a standard PCR protocol - rapid isolation of DNA and PCR assay for B-Globin, M.T. Vahey et al; enzymatic control of carryover contamination in PCR, J.L. Hartley et al; ultraviolet irradiation of surfaces to reduce PCR contamination, R.W. Cone et al; specificity, efficiency, and fidelity of the PCR, R.S. Cha et al; optimization and troubleshooting in PCR, K.H. Roux; long-distance PCR, O.S. Foord et al. Part 2 Sample preparation: rapid preparation of DNA for PCR amplification with gene releaser TM, E.P. Dawson et al; PCR amplification from paraffin-embedded tissues; sample preparation and the effects of fixation, C.E. Greer et al; RNA purification, J.J. Adamovicz et al. Part 3 Primer design: general concepts for PCR primer design, C.W. Dieffenbach et al; design and use of mismatched and degenerate primers, S. Kwok et al; mulitplex PCR, M.C. Edwards et al. Part 4 Detection of PCR products - quantitaion and analysis: immunological detection of PCR products, J.G. Lazar; quantitative PCR using the Amplisensor assay, C.N. Wang; DNA fingerprinting using arbitrarily primed PCR, M. McClelland et al; in situ PCR, G.J. Nuovu; single-strand conformational polymorphism, K. Fujita et al; genetic subtyping of human immunodeficiency virus using a heteroduplex mobility assay, E.L. Delwart et al; sensitive and fast mutation detection by solid-phase chemical cleavage, L.L. Hansen et al. Part 5 Starting from RNA: use of the PCR to quantitate relative differences in gene expression, W.C. Cause et al; quantitative liquid hybridization PCR method employing storage phosphor technology, M.T. Vahey et al; use of the SNuPE assay to quantitate allele-specific sequences differing by a single nucleotide, J. Singer-Sam; trapping internal and 3' -Terminal exons, P.E. Nisson et al; expression-PCR, D.E. Lanar et al. Part 6 PCR-mediated cloning. Part 7 PCR sequencing. Part 8 Cloning of PCR products. Part 9 Mutagenesis by PCR. Part 10 Alternative amplification technologies. Appendices: computer software for selecting primers; reagents and equipment. (Part contents).