In vivo cloning of PCR products in E.coli

Jonathan D. Oliner, Kenneth W. Kinzler, Bert Vogelstein

Research output: Contribution to journalArticle

Abstract

This report describes an efficient method to clone PCR products exploiting endogenous Escherichia colienzymatic activities. PCR products are engineered to contain terminal sequences identical to sequences at the two ends of a linearized vector. PCR products and vector DNA are then simply co-transfected into E. coli strain JC8679, obviating the requirement for enzymatic treatment of the PCR product or in vitro ligation. Thehigh rate of homologous recombination in this strain results in efficient incorporation of the insert into the vector, a process we refer to as invivo cloning(IVC).

Original languageEnglish (US)
Pages (from-to)5192-5197
Number of pages6
JournalNucleic acids research
Volume21
Issue number22
DOIs
StatePublished - Nov 11 1993

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ASJC Scopus subject areas

  • Genetics

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