PCR gene construction refs

ww40 William_D_WARREN at UMAIL.UMD.EDU
Wed Jul 20 15:38:31 EST 1994


I remember seeing a reference (I think it was in biotechniques) for using PCR
to combine DNA fragments in situations where you don't have convenient
restriction sites.

As I recall you design 4 oligos to amplify the two DNA pieces, with the oligos
were you want to join the fragments carrying sequences homologous to the other
product at their 5' ends. Amplify the two fragments separately then mix the
products of the two PCR reactions and use the outer primers only to amplify
the joined product.

Any refereneces for this much appreciated.

BILL

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William D. WARREN, PhD                    Email: ww40 at umail.umd.edu
Center for Agricultural Biotechnology     Phone: 1 (301) 405-7681
University of Maryland at College Park    FAX: 1 (301) 314-9075

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