Long PCR products

Dr. Duncan Clark Duncan at nospam.demon.co.uk
Tue Dec 1 11:51:32 EST 1998

In article <F3AK22.Msz.F at zoo.utoronto.ca>, Kowthar Salim
<kowthar at zoo.utoronto.ca> writes
>I am currently trying to amplify a region in a genomic DNA that is about 
>3kb long.

What type of genomic, bacterial, human, plant etc.?

3kb of bacterial should amplify with Taq. 3kb of human genomic will
present more of a problem and may require the use of a mix of Taq and
proof-reading pol. What are your exact PCR conditions i.e amount of
enzyme, no. cycles, amount of template etc.

>I have tried to increase the extension time (3 minutes) during the PCR 
>reaction.  Unfortunately, this has not yielded any PCR products.

3mins is fine for Taq or a pol. mix

>I was wondering if anyone has any suggestions or if any references can be 

Check out papers by Wayne Barnes in PNAS.


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

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