PCR cloning into T-vectors

ww40 William_D_WARREN at UMAIL.UMD.EDU
Tue Feb 18 11:18:00 EST 1992


Last year, mcgraw%gandal.dnet at SERVER.UGA.EDU wrote
>
>#1  Take a look at Marchuk et al (1991) NAR 19: 1154.  This is an easy way to 
>make a T-vector.  No restriction needed.  This works pretty well for us.
>
I have tried using this approach to clone PCR products and so far have had
no luck. I get a significant background of blues, suggesting that the T
addition isn't very efficient (at least for me) and around 10% whites (even
on no insert control ligations) and all whites that I've looked at so far
have been empty ! Can anyone give me an idea of the background usually
experienced when you clone in this way. If anyone out there buys the
commercial T-vector kits and does a no insert control ligation, do you get a
high background of white colonies ?

Any suggestions (at all) most welcome
BILL




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