PCR Cycle
cyc
cyc at glink.net.hk
Sun Mar 21 00:19:34 EST 1999
You may want to increase the extension time to 3-4 minutes as your target
site is 3000 bp. ie change the 72 degrees celsius segment from 1 minutes
to 3 minutes. This would give you enough time for the full 3000 bp to be
extended.
On Fri, 19 Mar 1999, Vellanoweth's Lab wrote:
> I am trying to amplify a gene that is 3000bp long using genomic plant
> DNA and I would like to know what the best times for each step in the
> cycle would be. The annealing temperature of the primers is 57oC and
> the current cycle I use is the following:
> 1. 95oC - 1 min
> 2. 40oC - 4 min
> 3. 72oC - 1 min
> 4. 95oC - 45 sec
> 5. 57oC - 2 min
> 6. 72oC - 1 min
> Then 34 cycles to step 4
********************************************************
Cheung Chung Yan,
Laboratory 511, Department of Microbiology,
The University of Hong Kong, Pokfulam Road, Hong Kong.
Telephone : (852) 28554379 Fax : (852) 28551241
¹q¡@¶l / email¡G cyc at hey.to
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