Temperature activity range for Taq-Ligase

Dr. Duncan Clark Duncan at nospam.demon.co.uk
Fri Jun 2 09:32:00 EST 2000


In article <3937AF87.D1FCC095 at imsb.au.dk>, Jesper <jsp at imsb.au.dk>
writes
>I am thinking about using Taq ligase in a cycling ligation but I was
>wondering wether it is active below 45 degrees celcius. I would like to
>use a denaturations step at 95 degrees then annealing at 20-30 degrees
>and ligation.... Is this possible with Taq ligase or is it only active
>at elevated temperatures (at the 45-65 stated in the New England Biolabs
>catalog).??

I would hazard a guess that yes it does work at the lower temps but that
the activity will be but a small percentage of the activity at the
higher temps. A bit like Taq polymerase in that respect. Could you not
anneal at the lower temp and then slowly ramp the temperature to say 50C
or so then denature and so on.

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

Duncan Clark
DNAmp Ltd.
Tel: +44(0)1252376288
FAX: +44(0)8701640382
http://www.dnamp.com
http://www.genesys.demon.co.uk






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