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. 1990 Sep 11;18(17):5107–5112. doi: 10.1093/nar/18.17.5107

Solid phase in vitro mutagenesis using plasmid DNA template.

T Hultman 1, M Murby 1, S Ståhl 1, E Hornes 1, M Uhlén 1
PMCID: PMC332130  PMID: 2205837

Abstract

Site-specific mutagenesis was accomplished using a solid support to generate single stranded vector and insert fragments which can be used to form gap-duplex plasmids through flanking, complementary double stranded regions. More than 80% mutants were obtained in both a single and a double primer approach. No special vectors or strains are needed and mismatch repair is avoided as the mutagenesis region is in a single stranded form when transformed into the Escherichia coli host cell. The fragments to be immobilized can be produced either by a polymerase chain reaction using general primers or by a site-specific restriction followed by a fill-in reaction. This novel method is rapid, simple and flexible and well suited for both manual and semi-automated in vitro mutagenesis protocols.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

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