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Gene. 2000 Feb 8;243(1-2):19-25.

Chain reaction cloning: a one-step method for directional ligation of multiple DNA fragments.

Author information

1
Wyeth Research, One Great Valley Parkway, Suite 30, Malvern, PA, USA. pachukc@labs.wyeth.com

Abstract

A novel DNA assembly method, chain reaction cloning (CRC), is described. CRC enables the ordered assembly of multiple DNA fragments in a single step. The power of the technique was demonstrated by the directed in vitro assembly of a plasmid comprised of six DNA fragments from a pool of 12 available fragments. The odds of obtaining the correct plasmid clone in a single step, using conventional techniques, is less than 1 in 191000000. Using CRC, the desired plasmid was recovered at a frequency of one in two. Ligation is no longer the rate limiting step in cloning, and limitless possibilities exist for the reconstruction of complex genomes.

PMID:
10675609
[Indexed for MEDLINE]

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