TA cloning

Dr. Duncan Clark duncan at nospam.demon.co.uk
Mon Jun 22 09:07:38 EST 1998


In article <6mk1kh$bma at mserv1.dl.ac.uk>, Wolfgang Schechinger
<wgschech at med.uni-tuebingen.de> writes
>Does this mean that Taq adds a dT under these conditions?

Yes.

>Will this work with Pwo or Pfu, too? (This would be the better 
>enzymes for cloning IMHO, since they have a lower error rate)

No. These enzymes have no terminal transferase activity.

Better still. Cut with EcoR V, tail with dTTP then ligate. Purify
unligated linear band from gel. Only that has the tail. This will
minimise the background tremendously.

Duncan


-- 
The problem with being on the cutting edge is that you occasionally get 
sliced from time to time....

Duncan Clark
DNAmp Ltd.
TEl/FAX 01252376288
http://www.dnamp.com
http://www.genesys.demon.co.uk



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