A highly efficient procedure for site-specific mutagenesis of full-length plasmids using Vent DNA polymerase.

Abstract:

:Careful titration of Vent polymerase activity allows efficient amplification of full-length plasmids (12 kb). The high processivity and fidelity of this enzyme made oligonucleotide-directed site-specific mutagenesis of plasmids a straight-forward process. Using only two primers, a mutagenic and a complementary, single-base mutants of recombinant plasmids were obtained consistently with > 90% efficiency from a single round of PCR. This procedure also made site-specific deletion, insertion, and several bases mutagenesis facile and efficient.

journal_name

Genome Res

journal_title

Genome research

authors

Byrappa S,Gavin DK,Gupta KC

doi

10.1101/gr.5.4.404

subject

Has Abstract

pub_date

1995-11-01 00:00:00

pages

404-7

issue

4

eissn

1088-9051

issn

1549-5469

journal_volume

5

pub_type

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