Amplitaq Gold in RapidCycler!

Dr. Duncan Clark Duncan at nospam.demon.co.uk
Thu Feb 17 09:45:05 EST 2000


In article <38abe135 at redeye.it.ki.se>, Csaba Kiss <csakis at ki.se> writes
>We are using a Rapid cycler from Idahotec. I used Platinum Taq polymerase to do 
>hot start PCR before and it works perfectly. Now we have a very good price offer 
>from PE to use Amplitaq Gold. If you want to use the Gold you have to keep your 
>sample at 95 degree for 10 minutes to activate the enzyme. 


>In the Rapid Cycler, 
>however, the denaturation step is very short, ususally 0 sec. Is it enough to 
>activate the enzyme fully, even if I insert a 10 minute 95 degree initial 
>incubation? 

The 95C ten minute step is a must. Without that you will get no
activation. After 10 mins approx. 50% of the total enzyme present is
active. With the speed of the Rapid Cycler and a 0 sec denaturation you
are unlikely to get any more enzyme activated. Using a normal PCR
machine one needs 30 plus cycles of say 30secs at 94C denaturation to
really get more released. You will also have to use the AmpliTaq Gold
reaction buffer. 

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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