PCR of A/T-rich regions

berschic at convex.rz.uni-duesseldorf.de berschic at convex.rz.uni-duesseldorf.de
Thu May 19 02:26:37 EST 1994


>I have to do a bunch of PCR on highly A/T rich sequences and was wondering
>if anyone has done any optimization experiments for any of the zillion PCR
>variables, as they relate to A/T sequences.  Lower extension temp? Higher salt?
>Increase gelatin?  Any thoughts on this would be appreciated.
>
>                       rcarper at jhuhyg.sph.jhu.edu
>
>                                    Thanks much
>
>                                        robin


Hi,
I didn´t try zillion optimization porocedures (we didn´t have so much
TAQ). What I did is using the annealing temperature also as extension temp.
This works very fine with an AMPFLP system Apo B. The only thing is you 
have to increase the annealing-extension time (up to 3-5 min for fragments
of about 1 kb) and perhaps increase the optimized annealing temp.
Hope this helps
peter
(berschic at convex.rz.uni-duesseldorf.de)



More information about the Methods mailing list