Long reads on ABI 373

Werner wwitkeNOwwSPAM at earthlink.net.invalid
Thu Mar 30 20:39:10 EST 2000


Anyone out there know how to get really long reads on an ABI
373?  We have slowly but surely been increasing the length
of
read on our 373 from 0 mismatches in 400 bases to 0
mismatches in 650 bases using the ABI pGEM control.  We have
tried things such as different polyacrylamide manufacturers,
thinner gels, lower percentage gels, membrane combs, less
wattage and higher PMT voltage.  We have not changed our
cycle sequencing protocols except that we use quarter
reactions. We have used the version 2.0 sequencing mix.  Our
error rate at 850 bases is about 1.5%.  This is about equal
to a 377.



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