Molecular cloning of complementary DNA: preparation of a plasmid vector with low transformation background |
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| Authors: | A Leriche D Christophe H Brocas G Vassart |
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| Affiliation: | Institute of Interdisciplinary Research, School of Medicine, Free University of Brussels, Campus Erasme, Bât. C, Route de Lennik 808, B-1070 Brussels, Belgium |
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| Abstract: | 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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| Keywords: | Chercheur qualifié at the Belgium “Fonds National de la Recherche Scientifique.” To whom correspondence should be addressed. |
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