Overview

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.

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