Protocol

Mutagenic PCR

This protocol was adapted from "Mutagenic PCR," contributed by R. Craig Cadwell and Gerald F. Joyce, Chapter 30, in PCR Primer, 2nd edition (eds. Dieffenbach and Dveksler). Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA, 2003.

The first 100 words of the full text of this article appear below.

INTRODUCTION

The top priority of mutagenic PCR is to introduce the various types of mutations in an unbiased fashion rather than to achieve a high overall level of amplification. Thirty cycles of PCR are carried out to ensure that mismatched termini have ample opportunity to become extended to produce complete copies. A large template input reduces the chance that any two molecules isolated from the final population will carry the same mutation. This protocol is derived from "standard" PCR conditions (Coen 1991) and makes a number of changes designed to enhance the mutation rate. The mutagenic PCR should be carried out . . . [Full Text of this Article]

MATERIALS

Reagents

Equipment

METHOD

TROUBLESHOOTING