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. 2023:2633:87-95.
doi: 10.1007/978-1-0716-3004-4_8.

In Vitro Site Directed Mutagenesis

Michael J McClellan  1
Affiliations

In Vitro Site Directed Mutagenesis

Michael J McClellan. Methods Mol Biol. 2023.

Abstract

Site-Directed Mutagenesis (SDM) allows for changes in the DNA sequence of plasmids using polymerase chain reaction (PCR). It is a reliable, accessible, and rapid method which is the common initial step of many biochemial or genetic experiments. Here we describe the various different forms of SDM before giving a detailed method for the introduction of substitutions, insertions, or deletions using a fast, ligation-free protocol, followed by colony PCR to screen for mutated sequences.

Keywords: Colony PCR; Genetic alteration; Mutation; Plasmid manipulation; Polymerase chain reaction; Site-directed mutagenesis.

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