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

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Cheung Chung Yan,
Laboratory 511,  Department of Microbiology,
The University of Hong Kong, Pokfulam Road, Hong Kong.
Telephone       : (852) 28554379  Fax   :  (852) 28551241

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