why gel is not running in straight line?? - (Nov/04/2009 )
Hello,
I have a problem with electrophoresis, my gels (regular cell lysates) are not running in a straight line, but thay are one side curved-up . does anyone have some idea why it is happening??
smily effect...
heat is not uniform across the gel...
try running at a lower voltage!!!
wat conditions are u running in???
is it just the dye showing this effect or the bands too???
dirty electrode.
Tilted gel tray with different buffer depths at each side.
phage434 on Nov 5 2009, 09:32 AM said:
or tilted gel tray when the gel is setting, which will lead to a gel of variable thickness.
Do you have a picture?
It usually works ok for me when I decrease to a lower voltage.
How much protien lysate per well?
this used to happen to me on the first and last well, I also cant figure it out why... maybe too high voltage is the culprit. My solution: i ignore the 1st and last well when i run my page gel...
Thanx everyone for your posts.
I usually run it at 70V at the beggining and 120-130 later, is it to high?? samples contain between 20-40 ug of protein in reduced condition.
porfirion on Nov 5 2009, 01:33 PM said:
I usually run it at 70V at the beggining and 120-130 later, is it to high?? samples contain between 20-40 ug of protein in reduced condition.
why dont u try running it at a constant voltage... instead of switching.. might help..
and 40 mcg is too much for a SDS-PAGE.. tat too iof u are silver staining it!!!