HELP sequencing PAC

Lars Borrmann schoko at
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?

  Thank you

  MA Pujana
  mpujana at

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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