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
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