PCR A Practical Approach

by McPherson, M. J.; Quirke, P.; Taylor, G. R.

• ISBN13: 9780199631964
• ISBN10: 0199631964
• Format: Paperback
• Copyright: 1991-10-10
• Publisher: IRL Press

Summary

The Polymerase Chain Reaction (PCR) has become one of the most widely used techniques in biomedical research, enabling the fast, inexpensive production of large quantities of DNA from minute amounts of starting material. Although the basic procedure is very simple, many variations of it havebeen developed for different applications, for use in such diverse areas as forensic and diagnostic medicine, genetic engineering, and the food industry.PCR: A Practical Approach provides a general introduction to PCR for researchers new to the area, followed by chapters detailing specific applications of the technique. It covers gene analysis at the level of restriction enzyme polymorphism, point mutation and sequence analysis. Methods for usingdifferent source materials - cloned DNA, genomic DNA, RNA, nucleic acid from archive material and PCR products themselves - are given.The emphasis throughout is on the practical applications of the technique and detailed experimental protocols are included at all stages, backed up by advice on avoiding potential pitfalls. This volume provides a starting point for those new to PCR and will also serve as a useful reference for moreexperienced researchers in all areas of biological science.

Table of Contents

Polymerase Chain Reaction: Basic Principles and Automation

Polymerase Chain Reaction in Genetic Diagnosis

Extraction of Nucleic Acid from Fresh and Archival Material

Analysis of Genomic Sequence Variation Using Amplification and Mismatch Detection and Direct Sequencing

Detection of Deletions and Point Mutations

PCR of TG Microsatellites

Genome Amplification Using Primers Directed to Interspersed Repetitive Sequences (IRS - PRC)

PCR Amplification of Microdissected DNA

Inverse Polymerase Chain Reaction

PCR-directed cDNA Libraries

PCR with Highly Redundant Primers

General Application of PCR to Gene Cloning and Manipulation

Semi-quantitative PCR for the Analysis of Gene Expression

The Fidelity of DNA Polymerases Used in the PCR

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