DNA SEQ- fade at 2000 scans- request for a revisit

Richard F. Cook (cook@MIT.EDU)
Thu, 19 Feb 98 11:44:51 EST

Dear DNA sequencing ABRFers,

We are having the infamous "fast fade" problem and in searching the
Automated DNA Sequencing WWW Page searchable index (many thanks to those
responsible for that fine page) I find varied and contradictory bits of
info.

Listed cures included: specific detergents, strong NaOH washes, strong HCl
washes, scalding hot water washes, using a dishwasher, voodoo and more.

My request is for anyone who thinks she or he really has a handle on this
problem, i.e. has managed to beat this problem for at least 6 months, to
please respond with your cure so I can make a better plan of attack.

PS - we have had some modest success with Multi-terge, at least we can read
the bases in most cases within the fade region, but still hope that there
is something better out there.

PPS - we did not have this problem 'till Big Dye came to town and we are
only doing 1:2 dilutions. Coincidence or not? The archives show fade
problems dating long before BD's but I'm wondering if larger dilutions of
DBs help the fade problem. Any chance of a lot problem here or is this a
permanent condition for everybody. We just saw the fade on a brand new set
of plates!

Thanks to all in advance,

Dick Cook