Apr 01, 2020
Site-directed mutagenesis
- 1Martin-Luther Universität Halle-Wittenberg
Protocol Citation: Johannes Stuttmann 2020. Site-directed mutagenesis. protocols.io https://dx.doi.org/10.17504/protocols.io.behgjb3w
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: April 01, 2020
Last Modified: April 01, 2020
Protocol Integer ID: 35080
Keywords: site-directed mutagenesis, mutagenesis, PCR, DpnI,
Abstract
Site-directed mutagenesis (SDM) as described here is used to introduce a mutation in the sequence of a plasmid of interest by PCR-based methods. Several different approaches have been previously reported, and also the generation of multiple mutations in a single step is possible. This protocol deals with the simplest version of SDM for production of a small mutation at a single position of a plasmid, e.g. for generation of a protein variant or elimination of a restriction site.
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