PCR A/T CLONING VECTOR

David R. Nelson dnelson at utmem1.utmem.edu
Tue Dec 20 10:07:34 EST 1994


In article <marcvdc.12.004438DC at lmb1.rug.ac.be>, marcvdc at lmb1.rug.ac.be
wrote:

> IS  IT POSSIBLE TO MAKE A PLASMID VECTOR WHICH GENERATES  SINGLE T-ENDS AFTER 
> DIGESTION WITH A CERTAIN  RESTRICTION ENZYME ? 
> 
> Such a plasmid would be very usefull for cloning PCR-fragments by  A/T 
> cloning.  
> MARC  VAN DE CRAEN
> 
Dear Marc,

	All you have to do is cut any vector with a blunt end cutter like EcoRV
and run the linearized plasmid through a couple cycles of PCR with only
dTTP present.  That will add on the one base overhang T that you want.

-- 
David R. Nelson
Assistant Professor
Dept. of Biochemistry
University of Tennessee, Memphis



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