A sad beginning but a good ending? - "Evolution" of tracking dye in SDS page (Apr/10/2009 )

Hi guys,

I think this is something weird.

Lately i been running SDS page, the tracking dye kinda diffuse in the beginning after going through the stacking gel . then in the mid way it sharpened up but still with some diffusion on the front . then in the end everything condensed up to a shart band looking good. but i thought everyhting supposed to condense up after passing the stacking gel.

The resolution of protein seems to be affected from this too.
I m suspecting something wrong with the buffer used to make the resolving gel

What do you guys think?

any hand on experience on this situation?

you could try a fresh lot of sds.

I agree with your suspicions -- probably something wrong with the buffer. If you prepared it yourself, make a new batch. If it's commercially prepared, call the company with the lot number and see if they have any info about problems with that lot.

There may be problem with the uniformity of the gel either due to buffer or polyacrylamide; or the gelling wasn't right. Try fresh buffer and polyacrylamide mix

hey guys

mystery solved.

Running buffer with a pH of 7.5 = Good game.

hai,is that mean that ur gel runs well after u change u running buffer wth ph 7.5?
can i ask what is the normal pH range for running buffer? thanks..