Home-Made TA cloning

Aydemir Akin akin at sage.cc.purdue.edu
Wed Jul 12 02:19:42 EST 1995

In article <3ttsks$6mf at gazette.bcm.tmc.edu>, Brian Desany
<bd033054 at condor.mbcr.bcm.tmc.edu> wrote:

> There are restriction enzymes that leave single base overhangs in the 
> middle of their site, XcmI (ccannnnn/nnnntgg) for example.  Two of these 
> sites with the appropriate bases chosen for the "n"'s should make it 
> really easy to TA clone - you don't have to ligate anything to the 
> cleaved vector before you do your actual TA cloning reaction.  Although 
> I haven't got around to trying it yet.


There is a published article on this.  You may want to take a look at it.
[Kovalic, D., et al.  General method for direct cloning of DNA fragments
generated by the polymerase chain reaction.  NAR 19 (16):4560, 1991.]

In their paper, they engineered pGEm5fZ(+) to have XcmI site. Digestion of
their vector with XcmI produces 'T' overhangs enabling PCR product cloning.

Aydemir Akin, D.V.M.

akin at sage.cc.purdue.edu

1175 Animal Disease Diagnostic Lab
Purdue University
West Lafayette, IN 47907-1175

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