need help-mysterious problem
p.pagel at gsf.de
Wed Dec 18 03:12:20 EST 2002
> I loaded for example 8 wells of the ten available.
> After this I stained the gel with Sypro Orange (and took a photo) in order
> to see if I have the same loading in each well (20 ug total proteins/ well).
> Each time the 2-3 central lanes are straight but the lateral lanes are
> curved toward the sides (convex look).
Your gel is smiling at you! ;-)
Two common reasons:
- differences in salt concentration among the different lanes or
just high salt concentration of all samples
- temperature differences along the gel
For the first problem try a different extraction buffer or dialyse the
salt out. It also helps to load the empty wells with a dummy sample
(extraction buffer + loading buffer).
If for some reason you have temperature problems you can try to run the
gel with a stirbar in the tank and put it on a stir-plate. Or keep
mixing the buffer with a pump.
Dr. Philipp Pagel Tel. +49-89-3187-3675
Institute for Bioinformatics / MIPS Fax. +49-89-3187-3585
GSF - National Research Center for Environment and Health
Ingolstaedter Landstrasse 1
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