Hi,
I was wondering if it is possible to modify the thermal cycling
conditions described in ABI protocols for Big Dye Termintors
The raccomended conditions are:
95°C for 30 sec
50°C for 15 sec
60°C for 4 min
In particular does anybody know if it is possible to shorten the
extension time?
If not, why, considered that in a PCR reaction the polymerase performs
about 1000-2000 bases /min? Is it because of a low extension temperature
(60°C)?
Does anyone has experimented succesfully other conditions?
========================
Barbara Simionati
CRIBI Biotechnology Centre
and Department of Biology
University of Padua, Italy
