HELP sequencing PAC

Lars Borrmann schoko at uni-bremen.de
Fri Apr 18 07:36:12 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?

  Thank you

  MA Pujana
  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)
Good luck




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