Samples not on a straight line caused by gel dryness? - (Dec/05/2008 )

Hi,
I'm still new in running gel electrophoresis.
After preparing resolving gel, I had to leave for a very long time (> 1 hour) while actually it only takes about 25 min to solidify. Afterwards, the samples ran with so different speeds that the sample "blue line" was twisted in an ugly way!
Could have the delay (slightly dry gel) caused the problem?
I also noticed that the thin metal wire of the electrophoresis gasket was not straight for some reason and the bubbles produced during electrophoresis process didn't rise up completely vertically. Could this be the reason? I've run at least 5 experiments so far and most of them give me similar pattern of distortion.
Thanks, Hanhan

Hanhan, if you pour your gels, you should put water or ethanol/isoprop on top of the resolving gel to omit drying. But nonetheless it does not sound to me, as if the gel is the source of your twisted sample run. It seems more likely, that you may have run your gel at a very high voltage and is may have get overheated. Then you get an smiley effect in your samples. Try to your your gels at a lower voltage or perform your electrophoresis in the cold room. Also fill in loading dye in every pocket, even in empty ones (only laemmli buffer w/o protein) to get an even running front.

Your airbubbles on the metal wire may go up lopsided, this is not bad, as long as there are bubbles, everything is o.k.

Thanks! I did pour isoprop on top. The smiley effect actually even started before I increased voltage from 80 to 130 v. In addition, I wrapped the tank with ice. So that's why I was really confused. Previously I got straight line. So it looks like there's sth. hidden somewhere and I'm not aware of. BTW, I was making full use of the 15-# (1 ladder plus 14 samples).

Hanhan, if you pour your gels, you should put water or ethanol/isoprop on top of the resolving gel to omit drying. But nonetheless it does not sound to me, as if the gel is the source of your twisted sample run. It seems more likely, that you may have run your gel at a very high voltage and is may have get overheated. Then you get an smiley effect in your samples. Try to your your gels at a lower voltage or perform your electrophoresis in the cold room. Also fill in loading dye in every pocket, even in empty ones (only laemmli buffer w/o protein) to get an even running front.

Your airbubbles on the metal wire may go up lopsided, this is not bad, as long as there are bubbles, everything is o.k.

Hm, strange. But why is it dry when you poured isoprop on top of it before leaving for 1h? And so it has had also enough time for polymerization. Can you try to use another electrophoresis module to test, if the problem lies within the apparatus or the gels?

I didn't test it using another module. I just suspected that the gels were a little bit dry at some edges, I didn't really pay much attention to them, probably not the reason. Thanks anyway, I'll try more.