PCR‐mediated Mutagenesis
J Chamberlain, University of Washington, Washington, USA
Published online: March 2004
DOI: 10.1038/npg.els.0003766
Abstract
PCR allows a defined sequence to be amplified with high specificity from complex mixtures of DNA. However, when starting with
cloned DNA as a template, the PCR can be performed under relaxed stringency conditions that allow short primers or primers
containing significant mismatches with the template to amplify the cloned DNA. These features can be used to alter the sequence
of a cloned template by the use of PCR primers with single or multiple base changes relative to the cloned template. Similarly,
primers can be designed that will either add or remove sequences from the final amplified product. In this manner a large
number of defined mutations can be introduced rapidly into cloned genes by PCR.
Keywords: recombinant PCR; primer; DNA polymerase; cloning
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