ABI 310 DNA sequencer and Pop6

Mick Guerini mguerini at gemini.oscs.montana.edu
Sun Mar 12 17:38:47 EST 2000


I have just discovered this resource and have two questions regarding
1)the application/use of ABI 310 DNA sequencers and 2)Pop6 sequencing
chemistry.

    Our department runs an ABI 310 using the long capillary 95% of the
time.   We primarily sequence cDNA library clones, genomic sequences,
mutagenesis templates and verify cloning inserts.  Because of our far
ranging applications we have decided to primarily  run the long
capillary.  What is this groups opinion on the ABI 310 and its ability
to function with the long capillary?  We are receiving conflicting
reports about running this instrument with the long capillary.  Does the
machine perform better with the short capillary?  We are afraid that we
are part of a very (I stress very) small group of users trying to
sequence long templates with the ABI 310.  Any thoughts on this subject
would be appreciated.

    Secondly, we have encountered in the last 60 days a problem relating
to the Pop6 chemistry.  Two lots in particular have problems and we have
determined this by trial and error and by finding a lot that worked
great for 105 runs (one whole bottle of polymer).  Let me explain the
problem briefly and see if anyone out there has any suggestions or
comments.  The problem is intermittent, 4 or 5 runs may work well then a
run has good peaks (crisp, tight, tall) for the first 200 some odd bases
and then the sequence loses resolution and looks like wide waves. This
problem is intermittent in the first 10-12 runs on a new capillary and
then the capillary goes bad.  Does anyone have this problem with Pop6
currently (or previously)?  I can tell you that our diagnostics of the
machine reports no problems ( I ran them myself).  We have put in a new
syringe, made sure the syringe step mechanism is correctly calibrated,
installed all new ferrels, a new pump block and we use injection grade
water in our machine (make sure we have no/few impurities).  This
problem happens regardless of the number of capillaries we have
installed  (currently 11 different capillaries from three different
manufacturing dates have all had the same problem).

    An additional question:  Has new chemistry been developed for the
3700 (96 capillary machine) and does it work in the 310?

    Any feedback on the ABI 310, expereinces using the instrument or
problematic Pop6 would be appreciated.  We are trying to find the right
combination for running our machine correctly.

    Thanks for your time,


    Mick Guerini
    Department of Veterinary Molecular Biology
    Marsh Laboratories
    Montana State University
    Bozeman,  MT    59717

    mguerini at gemini.oscs.montana.edu

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