SDS PAGE - Bubble inside the Gel (Nov/20/2010 )

Dear Friends,

I made a SDS-PAGE gel to run my protein samples. When I prepare the seperating gel and overlay the 70% ethanol, the gel was in a good condition. Then I made the stacking gel and placed the comb. After 30 minutes I found some bubbles(~1mm dia) in the middle of the seperating gel.

Could anyone advice me in this regard. How can I get rid of this issue in future?

Thanks in advance!

With Regards,
Selvam Ayarpadikannan.

-biocrazy-

Did you degas the solutions?

-hobglobin-

hobglobin on Sat Nov 20 14:08:45 2010 said:

Did you degas the solutions?

Dear Hobglobin,

Usually we dont degas the solutions. Could you tell me how to degass the solutions.........

-biocrazy-

Very few people ever degas their solutions for gel preparation. So, in my opinion this is not an issue. We have had this problem as well. One issue is if you didn't let the separating gel polymerize well before adding the concentrating gel. Otherwise I have seen this happen when people removed the comb out of the gel. I remove the comb when the gel is in the electrophoresis chamber.

-BioMiha-

this could also happen if the gel is poured with cold solutions. trapped gasses will be released and form bubbles. if you allow the solutions to reach room temperature before pouring then degassing will be unnecessary.

-mdfenko-

mdfenko on Mon Nov 22 20:34:25 2010 said:

Dear Biomiha and mdfenko,

Thanks a lot for your valuable advices!

-biocrazy-