your gels look a litte like our "bizzare gels"
(http://www.mpiz-koeln.mpg.de/~weisshaa/Adis/Bizzare-Gels.html). After so=
me
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 t=
he
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 ca=
n
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@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
Fax: +49-221-5062 313
e-mail: weisshaa@mpiz-koeln.mpg.de
WWW-URL: http://www.mpiz-koeln.mpg.de/~weisshaa/BW-info.html