PCR A/T CLONING VECTOR
Richard T. Timmer
rttimme at unix.cc.emory.edu
Tue Dec 20 18:24:36 EST 1994
Michael Cooley (szcooley at dale.ucdavis.edu) wrote:
: 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. The advantage would be that you only need to digest the vector to be
: : certain that all the 3' ends contain only one T .
: : OR DOES ANYONE KNOW WHERE I CAN BUY SUCH A PLASMID ?
: : I realise that clonetech and promega both offer PCR cloning kits but they only
: : offer linearised plasmids generated by ddTTP tailing of blunt end plasmids.
: : Thank you in advance for your nice reply
: : MARC VAN DE CRAEN
: : E-MAIL: MARCVDC at LMB1.RUG.AC.BE
: I was sent a protocol for making a T/A cloning vector using the TAQ
: polymerase to add the necessary T's to the end of a blunt cut vector. I
: haven't tried it as yet, but considering the ridiculous cost of
: Invitrogen's plasmid I am likely to try it soon. YOU HEAR THAT INVITROGEN!!!
Could you post your protocol for generating the T/A cloning vector using
Dept. of Physiology
Atlanta GA 30322-3110
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