This PCR manual seeks to provide a solid grounding in basic techniques in addition to clinical diagnostic applications. Detailed protocols are given for the use of PCR research and clinical laboratories, backed by information at each step. This second edition includes several new aspects, such as: how to set up a PCR lab and how to control contamination and false negative results; isolation of RNA from clinical samples; sequencing of PCR products; long range PCR; nucleotide analogues in PCR; PCR cloning, RT-RNA PCR, microsatellite PCR, in situ PCR; and single-cell cDNA PCR and application to forensic analysis.