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L. v.d. Zande - cloning PCR fragments

Aida Metzenberg aida at CGL.UCSF.EDU
Thu Oct 29 18:24:44 EST 1992

No method is perfect, but our current favorite is to use Bernie
Weisblum's pDK101 vector.  It has a special linker in the Nco I
site that when digested with XcmI leaves 1-base T overhangs.  You
can directly clone fragments from Taq Pol amplifications, with an
efficiency advantage over blunt cloning.  The reference is

 Kovalic D; Kwak JH; Weisblum B.
     General method for direct cloning of DNA fragments generated by the
     polymerase chain reaction.
   Nucleic Acids Research, 1991 Aug 25, 19(16):4560.
     (UI:  91360369)

I agree with Dr. Bloksberg, though, that large fragments can be more of a pain.

Aida Metzenberg
aida at cgl.ucsf.edu

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