UNIT 8.1 Oligonucleotide-Directed Mutagenesis without Phenotypic Selection

  1. Thomas A. Kunkel

Published Online: 1 MAY 2001

DOI: 10.1002/0471142727.mb0801s13

Current Protocols in Molecular Biology

Current Protocols in Molecular Biology

How to Cite

Kunkel, T. A. 2001. Oligonucleotide-Directed Mutagenesis without Phenotypic Selection. Current Protocols in Molecular Biology. 13:8.1:8.1.1–8.1.6.

Author Information

  1. National Institute of Environmental Health Sciences, Research Triangle Park, North Carolina

Publication History

  1. Published Online: 1 MAY 2001
  2. Published Print: JAN 1991


A DNA sequence can be specifically altered by synthesizing the desired sequence change within an oligonucleotide, and then converting this into a biologically active circular DNA strand by using the oligonucleotide to prime in vitro synthesis on a single-stranded circular template. This protocol uses a DNA template containing a small number of uracil residues in place of thymine. Use of the uracil-containing template allows rapid and efficient recovery of mutants; in principle this same template can be applied to most of the other mutagenesis protocols in use.