Unit

UNIT 10.2 Overview of PCR

  1. Christine D. Kuslich1,
  2. Buena Chui2,
  3. Carl T. Yamashiro3

Published Online: 1 OCT 2008

DOI: 10.1002/9780470089941.et1002s00

Current Protocols Essential Laboratory Techniques

Current Protocols Essential Laboratory Techniques

How to Cite

Kuslich, C. D., Chui, B. and Yamashiro, C. T. 2008. Overview of PCR. Current Protocols Essential Laboratory Techniques. 00:10.2:10.2.1–10.2.31.

Author Information

  1. 1

    Molecular Profiling Institute, Phoenix, Arizona

  2. 2

    GE Healthcare, Piscataway, New Jersey

  3. 3

    Arizona State University, Tempe, Arizona

Publication History

  1. Published Online: 1 OCT 2008
  2. Published Print: JAN 2008

Abstract

As a means of rapidly copying and amplifying a selected template sequence from a pool of DNA in vitro, the polymerase chain reaction (PCR) as a stand-alone technique and in combinations with other methods has a vast range of applications. This chapter provides an overview of the theory and applications for this powerful and versatile laboratory method. A generic protocol for the broadest application is described in this unit along with the basic theory underpinning PCR to foster an understanding of how to make modifications to the protocol that can be applied to specific applications of the PCR technique. There is a troubleshooting table provided to describe and resolve the most common problems associated with PCR, as well as a table for suppliers and manufacturers of thermal cycling instruments. A detailed discussion of various DNA polymerases and suppliers is also provided.

Keywords:

  • polymerase chain reaction (PCR);
  • dNTPs;
  • thermal cycler;
  • DNA polymerase;
  • DNA template;
  • denaturation;
  • annealment;
  • elongation;
  • melting temperature (Tm);
  • hot start;
  • primer design;
  • contamination