1. An improved PCR-mutagenesis strategy for two-site mutagenesis or sequence swapping between related genes.
- Author
-
Kirsch RD and Joly E
- Subjects
- Amino Acid Substitution, Animals, Base Sequence, COS Cells, Chromatography, High Pressure Liquid, DNA chemistry, DNA metabolism, DNA Primers chemistry, Oligodeoxyribonucleotides chemistry, Oligodeoxyribonucleotides isolation & purification, Plasmids, Point Mutation, Reproducibility of Results, Transfection, Mutagenesis, Site-Directed, Polymerase Chain Reaction methods
- Abstract
The QuikChangeTM protocol is one of the simplest and fastest methods for site-directed mutagenesis, but introduces mutations at only one site at a time, and requires two HPLC-purified complementary oligonucleotides. Here, we describe that this method can be used with non-overlapping oligonucleotides. By doing this, two separate sites can be mutagenised simultaneously, or money can be saved by using a second 'standard' oligonucleotide. By a further modification, we have also used the QuikChangeTM approach to exchange DNA sequences between closely related genes.
- Published
- 1998
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