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Nucleic Acids Research, 1993, Vol. 21, No. 22 5192-5197
© 1993

METHODS

In vivo cloning of PCR products in E.coli

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

The Johns Hopkins Oncology Center Baltimore, MD 21231, USA

*To whom correspondence should be addressed

Received July 12, 1993. Revised September 30, 1993. Accepted September 30, 1993.

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).


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