UNIT 8.5 Directed Mutagenesis Using 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
Cormack, B. 2001. Directed Mutagenesis Using the Polymerase Chain Reaction. Current Protocols in Molecular Biology. 37:8.5:8.5.1–8.5.10.
- Published Online: 1 MAY 2001
- Published Print: JAN 1997
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.