T-tailing for PCR product ligation

G Selvarani medp3056 at leonis.nus.sg
Mon Sep 19 06:36:30 EST 1994


Loryn Sellner (lsellner at uniwa.uwa.edu.au) wrote:

: Dear netters,
: I have been reading the advice for cloning of PCR products by using the 
: TA cloning vector (or any T tailed vector). Is there a way to T tail my 
: own vector so I can clone my PCR product into the site I want, (namely a 
: blunt ended restriction site)?
: Thanks.
: Loryn

dear loryn, 
you can prepare your own ta vector using preferably a vector containing 
Eco RV site.after the blunt ending,incubate the restricted dna with Taq 
polymerase and dTTP at 72oC for about 2 hours and clean them as normal dna.
there was an article regarding this procedure in Nature of either May,Jun 
or July 1994.If you can't locate it E-mail me and i'lltry to search for you.
I've tried it and it works as good as the commercial ones.
Happy trying!


arun




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