Abstract
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.
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