Site-directed, Ligase-Independent Mutagenesis (SLIM) for highly efficient mutagenesis of plasmids greater than 8kb

被引：68

作者：

Chiu, Joyce ^{[1
]}

Tillett, Daniel ^{[2
]}

Dawes, Ian W. ^{[1
]}

March, Paul E. ^{[3
,4
]}

机构：

[1] Univ New S Wales, Sch Biotechnol & Biomol Sci, Sydney, NSW 2052, Australia

[2] La Trobe Univ, Sch Pharm & Appl Sci, Bendigo, Vic 3550, Australia

[3] Univ Cambridge Emmanuel Coll, Dept Chem, Boston, MA USA

[4] Univ Cambridge Emmanuel Coll, Dept Biol, Boston, MA USA

来源：

JOURNAL OF MICROBIOLOGICAL METHODS | 2008年 / 73卷 / 02期

关键词：

site-directed mutagenesis; inverse PCR; ligase-free; 5'-adapter;

D O I：

10.1016/j.mimet.2008.02.013

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Modifying the Site-directed, Ligase-Independent Mutagenesis (SLIM) protocol from a single reaction mode to a two-reaction mode enables highly efficient mutagenesis of plasmid constructs that exceed 8kb. This modified approach reduces the complexity of the PCR step and is optimised for generation of heteroduplexes from long PCR products. The two-reaction mode SLIM has 92% efficiency. (c) 2008 Elsevier B.V. All rights reserved.

引用

页码：195 / 198

页数：4

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