Promega T-vector

nsaunders at molbiol.ox.ac.uk nsaunders at molbiol.ox.ac.uk
Wed Sep 11 14:33:59 EST 1996


I'd agree with the last post in praise of the Promega T-vector. I've used this 
several times and it only failed me once, when I had a genuinely un-clonable 
product that no other method could handle either.
If I have a good yield of clean, specific PCR product, I generally ligate 1 
microlitre of T-vector and 5 or so microlitres of PCR product in 10 
microlitres, o/n at 4C, then use it all in a transformation. Sometimes 
transformation efficiency is quite low and there are usually a few blues, but 
there are always enough whites to work with, most of which have the insert. 
For small fragments there are often pale blues with the insert too, as 
detailed by Promega. The other trick they suggest is to harvest the cells 
after the LB incubation and resuspend in a smaller volume, which also works 
well.

Neil Saunders

Dept of Biochemistry,
University of Oxford

(no affiliation etc)




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