PCR A/T CLONING VECTOR
szcooley at dale.ucdavis.edu
Mon Dec 19 20:18:29 EST 1994
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!!!
More information about the Methods