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Molecular cloning of complementary DNA: Preparation of a plasmid vector with low transformation background

✍ Scribed by Anne Leriche; Daniel Christophe; Huguette Brocas; Gilbert Vassart

Book ID: 102629285
Publisher: Elsevier Science
Year: 1983
Tongue: English
Weight: 281 KB
Volume: 129
Category: Article
ISSN: 0003-2697
DOI: 10.1016/0003-2697(83)90077-5

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✦ Synopsis

A simple method that allows the rapid preparation of oligo dG-tailed plasmid vectors is presented. The procedure involves purification of the tailed molecules by hybridization to oligo dC-cellulose followed by a stepwise thermal elution. The resulting plasmid is virtually devoid of transformation activity in the absence of oligo dC-tailed DNA fragments. It allows construction of cDNA libraries with as low as 1% of colonies harboring wild-type plasmids.

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