t-tailing vector of choice

m-brazil at nimr.mrc.ac.uk m-brazil at nimr.mrc.ac.uk
Mon Sep 12 07:40:07 EST 1994

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> From: lsellner at uniwa.uwa.edu.au (Loryn Sellner)
> Reply-To: lsellner at uniwa.uwa.edu.au (Loryn Sellner)
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> Subject: T-tailing for PCR product ligation
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> 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 do that very easily. Digest your vector of choice with a
blunt cutter, and if possible heat denature the enzyme. Then incubate in a
larger volume with Taq polymerase and dTTP. 

I use this method (Marchuk et al, NAR 19, 5 , 1154) and it works well.

Hope it works for you

(m-brazil at uk.ac.mrc.nimr)

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