Dear George,
your gels look a litte like our "bizzare gels"
(http://www.mpiz-koeln.mpg.de/~weisshaa/Adis/Bizzare-Gels.html). After some
discussion on the ABRF list and several trys to get rid of the problem we
are now sure that the problem is related to a substance which sticks to the
glas plates. We got good results again after we changed the paper to dry
the gels and left out isoprop during cleaning.
To confirm our finding, last week a person came back to the lab who had
been trained some time ago to do fragment analysis. He used isoprop
together with the old paper batch - and the problem re-appeared.
Initially, I could not belive the story. But it really seems as if one can
deposit a quenching component on the plates which runs in the gel and
either absorbs the blue laser or the emission of the dyes.
Best regards, Bernd
>Greetings,
>>Has anyone seen gels where the samples start off looking like they all
>failed priming, then they suddenly "appear" and grow in strength? My
>first thought is that we lost the small fragments from the sequencing
>reaction in the clean-up process. However, the other half of the
>reaction plate looks fine (we do 96-well plates loaded across two
>48-well machines). This seems like it might be localized to a couple of
>plate sets, but it's hard to tell.
>>We've just begun using RapidRanger and LongRanger 1X premixes, but this
>might be a coincidence, since this problem has only affected three gels
>out of a hundred so far.
>>Instead of dirtying up the bandwidth, I've included a link to a page
>showing the gel snapshots.
>><http://www.genetics.wisc.edu/staff/mayhew/oddgels.html>
>>Any thoughts would be helpful.
>>Thanks.
>>--
>"People are DNA's way of making more DNA."(Edward O. Wilson, 1975)
>\ / \ \ / \ \ / \ \ George F. Mayhew / \ \ / \ \ / \ \ /
> \ /\ \ /\ \ /\Genome Center of Wisconsin /\ \ /\ \ /\ \
> _\/ \__\/ \__\/<mailto:mayhew at genetics.wisc.edu>_\/ \__\/ \__\
_____________________________________________________________________
PD Dr. Bernd Weisshaar
MPI fuer Zuechtungsforschung
Carl-von-Linn=E9-Weg 10
50829 Koeln
Germany
Tel. office: +49-221-5062 590 Tel. lab: +49-221-5062 311
=46ax: +49-221-5062 313
e-mail: weisshaa at mpiz-koeln.mpg.de
WWW-URL: http://www.mpiz-koeln.mpg.de/~weisshaa/BW-info.html
