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T/A PCR cloning

Graham Clark graham.clark at lshtm.ac.uk
Mon Dec 2 05:29:26 EST 1996


Iain Johnstone asks:

>Does anyone have a method for adding a single T to the OH end of a blunt
>cut plasmid to permit cloning of Tac amplified PCR products directly.  I
>assume it is possible using a dideoxy T and terminal transferase,
>however an actual method that is known to work would be useful.

Just such a method was published in Nucleic Acids Research (vol 19 (5),
p 1156) in 1991 by Holton and Graham. I have not tried it myself however.

Graham Clark



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