T/A PCR cloning

Holger Bohnemeier bohnemeier at ukbf.fu-berlin.de
Wed Dec 11 04:44:28 EST 1996


In article <329D948E.776D at udcf.gla.ac.uk>, gbga14 at udcf.gla.ac.uk says...
>
>Does anyone have a method for adding a single T to the 3'OH of blunt cut 
>plasmid for use in A/T Tac generated PCR fragment cloning.  I am working 
>on the assumption that dideoxy T and terminal transferase are used, 
>however a protocol known to work would be very useful

I have an article from BioTechniques 20:20-22 (1996) written by N. Hadjeb
and G.A. Berkowitz: Preparation of T-Overhang Vectors with High PCR Product
Cloning Efficiency.
It may point out exactly your interests.

Sincerely

-- 
                               Holger Bohnemeier
___           ___     Department of Clinical Pharmacology
\  \ +-----+ /  /    University Hospital Benjamin Franklin
 \  \| O O |/  /          Free University of Berlin
  |           |               Hindenburgdamm 30
 /  /| O O |\  \             12200 Berlin GERMANY
/__/ +-----+ \__\  phon: +49-30-8445-2289, fax: +49-30-8445-4482
                       e-mail: bohnemeier at ukbf.fu-berlin.de




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