ABI 373A Read Length Improvements

[Mick Jones]

Hi Netters

We have a 373A ABI DNA sequencer with 24 cm well to read plates (I know we are dinosaurs with no 
Stretch upgrade).  We routinely run 6% PAGE/Urea gels, and can read out to beyond 500 bases with 
good templates and good cycle sequencing reactions.  Although we are happy with our sequence 
data I was wondering if it could be improved.

My question is can we improve the length of read, accuracy, peak resolution by modifying the % 
of the PAGE.  Would we get an improvement by say running 5% gels (or even 4.5% gels)?  Would 
increasing run time from 12 to 14 or 16 hours coupled with 5% gels give better results?  
Would FMC Long Ranger be any benefit to us?

If anybody out there has a preferred set of running conditions then I'd be pleased to heare from 
you (gel concentration, TBE receipe, gel running conditions [watts, temp, current, etc]).  All 
would be appreciated, as I would hate to re-invent the wheel on this one.

Thanks to one and all in advance.

Mick Jones
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Mick Jones                      Tel: 081-740-3328 (+44-81-740-3328)
Department of Virology          FAX: 081-743-8331 (+44-81-743-8331)
RPMS,  Du Cane Road              Email: mjones at rpms.ac.uk 
London,  W12 0NN,  UK          URL: http://www.rpms.ac.uk/virology/virolhome.html
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