HELP sequencing PAC
schoko at uni-bremen.de
Fri Apr 18 07:41:53 EST 1997
Mara Dierssen wrote:
We are trying to sequence the ends of PACs (phenol/clorf purified)
with SP6 and T7 primers but we are not succesfully.
Does anybody know to do it?
mpujana at iro.es
We tried to sequence different PAC-clones several times but mostly we
had no results or bad results with lots of unknown nucleotides. So we
talked with a sequencing company who confirmed that sequencing of PAC
clones is very difficult. For better results we are now purifying
PAC-DNA with QIAGEN columns with a slightly modified protocol and
resuspend the pellet in bidest. For sequencing we use extended SP6 and
T7 primers (extend up to the BamH1 cloning site)
More information about the Methods