Unit

UNIT 8.5 Directed Mutagenesis Using the Polymerase Chain Reaction

  1. Brendan Cormack

Published Online: 1 MAY 2001

DOI: 10.1002/0471142727.mb0805s37

Current Protocols in Molecular Biology

Current Protocols in Molecular Biology

How to Cite

Cormack, B. 2001. Directed Mutagenesis Using the Polymerase Chain Reaction. Current Protocols in Molecular Biology. 37:8.5:8.5.1–8.5.10.

Author Information

  1. Harvard Medical School, Boston, Massachusetts

Publication History

  1. Published Online: 1 MAY 2001
  2. Published Print: JAN 1997

Abstract

This unit contains two basic protocols for introducing base changes into specific DNA sequences. The first describes the incorporation of a restriction site and the second details the generation of specific point mutations. An Alternate Protocol describes generating point mutations by sequential PCR steps. Although the general procedure is the same in all three protocols, there are differences in the design of the synthetic oligonucleotide primers and in the subsequent cloning and analyses of the amplified fragments.