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Peak stretching around 200bp

charlie charlie
Fri Feb 7 09:50:10 EST 1997


In article <5cnrkv$icn at net.bio.net>, Dr Tim Sawbridge,
T.Sawbridge at forbio.com.au wrote:

> Dear Sequencers,
>                 we have been seeing stretching of peaks from around 190
> to 210 bp across the gel on a 377 with resulting loss of resolution. The
> peaks return to their normal width after this so it seems to be a
> localised problem at around these fragment sizes. We are doing 3.5 hour
> runs on 36cm plates using dye-terminator chemistry with the ABI kit on
> double stranded plasmid DNA. Has anyone seen this and know what might be
> causing it? 
> Thanks in advance
> Tim Sawbridge
> 
> T.Sawbridge at forbio.com.au


We have been having the exact same problem (377, DT, 36cm, 4X run). We
thought it was a problem in the Gel reagents but it continued after
changing them. We acid cleaned the plates but that didn't help.It happens
on both a 4% polyacrilimide gel and on the Long Ranger gels. We usually
pour our gels in the evening and run them the next morning. Maybe these
clues will help. Please respond with some help if you figure this out.

Thanks, Charlie Demma   Demma at welchlink.welch.jhu.edu



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