Home-Made TA cloning
Srinivas K. Janardan, M.D.
vjanarda at umich.edu
Thu Jul 13 09:59:12 EST 1995
In article <3ttsks$6mf at gazette.bcm.tmc.edu>, Brian Desany
<bd033054 at condor.mbcr.bcm.tmc.edu> wrote:
In our lab, I developed a plasmid from pBluescript that has an additional
30+ bases in it at the MCS that contains two XCM I sites. The additional
insert is designed to yeild two T single base overhangs after XCM I
digest. This is then ready to use for TA cloning. I tried it a few times
but it did not appear to work that well. Right now we are using the
invitrogen kit. Someday I might pull this plasmid out of the freezer and
trry it again. BTW I have no connection to invitrogen.
> 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.
More information about the Methods