IUBio

Gel Problems

Bernd Weisshaar weisshaa at mpiz-koeln.mpg.de
Thu Feb 4 14:47:41 EST 1999


Dear Peter,

although not as strong, we had similar problems on our 377/96 sequencing
gels after switching from EtOH precipitation to G50 Sephadex purification
of probes. The solution was to wash the column material with 5 volumes of
water prior to using it (we have the Sephadex swelling in the microtiter
plates). After that, the column-purified probes run just great.

Best regards, Bernd


>We have recently experienced gels where adjacent lanes merge or overlap
>into each other.  This occurs to varying degrees ranging from a slight
>overlap of the lanes to a complete merging where the lanes become
>indistinguishable.  For 36 lane gels (seen in only a couple of
>instances) the degree on merging remains consistent throughout the
>entire read length of the gel.  In 96 lane gels (seen in about 50% of
>cases) the merging is severe at the low molecular weight region of the
>gel (0-200 bases) but then the lanes diverge as the run proceeds.  This
>is definitely not a lane-to-lane leakage issue.
>
>We are using the GeneScan application of the PE Biosystems 377 and are
>using Long Ranger Singels acrylamide.
>
>Has anyone else seen this phenomena before or are there any suggestions
>/ theories for causes / cures etc.
>
>Peter Johnson
>UK Forensic Science Service

_____________________________________________________________________

PD Dr. Bernd Weisshaar
MPI fuer Zuechtungsforschung
Carl-von-Linné-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 at mpiz-koeln.mpg.de
WWW-URL: http://www.mpiz-koeln.mpg.de/~weisshaa/BW-info.html




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