capillary sequencing

Peadar O'Gaora pog30 at sm.ic.ac.uk
Wed Feb 21 08:07:39 EST 1996


Dear sequencers,

We have recently had a capillary electrophoresis system for automated
sequencing installed by ABI.  It's the Prism CE310.  So far we've had
mixed results using it.  When it works it's beautiful, but we get quite a
lot of failures because of low peak values.  The amount of template isn't
the problem - that has been checked on gels.  Primer etc. seem to be fine
so we think its some subtlety in the template preparation.  We've been
mostly using Qiagen spin preps which don't work unless we precipitate them
and even then it's very variable.  Using the Tip-20 or Tip-100 preps are
fine but since we want to put quite a lot of templates through it's
financially and logistically less practical.  Does anybody have a
preferred template prep that works with this system - I forgot to mention
that we're using Taq FS enzyme with in dye terminator reactions.  If so
I'd love to hear about it and be forever in your debt because this is
driving me crazy at the moment.

many thanks,

Peadar



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