A method for multi-site-directed mutagenesis based on homologous recombination

被引：19

作者：

Liang, Xiquan ^{[1
]}

Peng, Lansha ^{[1
]}

Li, Ke ^{[1
]}

Peterson, Todd ^{[1
]}

Katzen, Federico ^{[1
]}

机构：

[1] Life Technol, Carlsbad, CA 92008 USA

来源：

ANALYTICAL BIOCHEMISTRY | 2012年 / 427卷 / 01期

关键词：

Mutagenize; Oligonucleotide; Plasmid; Mutagenic; Primer; PCR; ESCHERICHIA-COLI; DNA; RESTRICTION;

D O I：

10.1016/j.ab.2012.05.002

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

We have developed an efficient method for the simultaneous introduction of up to three mutations in a plasmid DNA via homologous recombination. The strategy is compatible with a variety of mutations, including degenerate codons in plasmids of different sizes. In contrast to other methodologies, this approach employs the same set of reagents for both single- and multi-site mutagenesis assays, minimizes the required protocol steps, and exhibits remarkably high mutagenesis efficiencies. (C) 2012 Elsevier Inc. All rights reserved.

引用

页码：99 / 101

页数：3

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