Unit

UNIT 3.17 Constructing Recombinant DNA Molecules by PCR

  1. Elaine A. Elion,
  2. Pablo Marina,
  3. Lu Yu

Published Online: 1 APR 2007

DOI: 10.1002/0471142727.mb0317s78

Current Protocols in Molecular Biology

Current Protocols in Molecular Biology

How to Cite

Elion, E. A., Marina, P. and Yu, L. 2007. Constructing Recombinant DNA Molecules by PCR. Current Protocols in Molecular Biology. 78:IV:3.17:3.17.1–3.17.12.

Author Information

  1. Harvard Medical School, Boston, Massachusetts

Publication History

  1. Published Online: 1 APR 2007
  2. Published Print: APR 2007

Abstract

This unit describes the use of PCR to construct hybrid DNA molecules. The unit provides an overview of how PCR can be exploited to accomplish numerous cloning strategies. The Basic Protocol outlines the PCR amplification and cloning strategies. The Commentary includes a troubleshooting guide for problems most frequently encountered in PCR cloning, plus four specific examples of the application of this technique to create in-frame fusion proteins, to create recombinant DNA products, to generate deletions and inversions by inverse PCR, and to introduce mutagenized PCR products or specific mutations or fusions by gap repair in yeast.

Keywords:

  • PCR;
  • recombinant DNA;
  • insertions;
  • deletions;
  • fusion proteins