Peak stretching around 200bp

Harold Harold
Wed Jan 29 14:25:41 EST 1997


>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

Tim,

We found the solution of this problem is to use taurine buffer for
preparing and running the gel.  This is know as glycerol tolerant buffer.
Lauara Livingstone at UNC-Chapel hill is the one who discovered this
solution.  We had it on our 377's and fortunately Laura rescued us.  ABI
did not have a solution.

I hope you will be able to open the attachment.

Hal


Harold G. Hills, Ph.D. DNA Sequencing Specialist	515 294-9585
1184 Molecular Biology Building			    FAX 515 294-1597
Iowa State University
Ames, IA 50011-3260				  hhills at iastate.edu
http://biotech.zool.iastate.edu/Biotech/Facilities/DSSF/homepage.html




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