UNIT 3.17 Constructing Recombinant DNA Molecules by the Polymerase Chain Reaction
Published Online: 1 MAY 2001
Copyright © 2003 by John Wiley and Sons, Inc.
Lab Protocol Title
Current Protocols in Molecular Biology
How to Cite
Elion, E. A. 2001. Constructing Recombinant DNA Molecules by the Polymerase Chain Reaction. Current Protocols in Molecular Biology. 3:IV:3.17.
- Published Online: 1 MAY 2001
- Published Print: OCT 1993
This is not the most recent version of the article. View current version (1 APR 2007)
This unit describes the use of PCR to construct hybrid DNA molecules. The main objective is to give an overview of how PCR can be exploited to accomplish numerous cloning strategies; it is assumed that the reader is already familiar with basic molecular biology techniques including PCR amplification and subcloning. The Basic Protocol outlines the PCR amplification and cloning strategies. A troubleshooting guide for problems most frequently encountered in PCR cloning, and three specific examples of this technique--for creating (1) in-frame fusion proteins, (2) recombinant DNA products, and (3) deletions and inversions by inverse PCR--are also presented.