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making gel in advance - to save time (Nov/06/2010 )

Hi

Has anyone prepared SDS page gel for western 24hr before? Is it possible to set the get, remove the combs, and load it inside the tank with running buffer overnight to run the gel the following morning?

-SF_HK-

We usually make them well in advance. I have run months old gels with no problems. You just can't let them dry. We usually just put them in deionized water until use, or you can put it in the tray with the running buffer. No worries.

-BioMiha-

Don't remove comb and wrap gel in paper towels moisturised with deionized water. Put in plastic bag and store in 4degree.

-ksturm-

ksturm on Sat Nov 6 23:33:39 2010 said:


Don't remove comb and wrap gel in paper towels moisturised with deionized water. Put in plastic bag and store in 4degree.


Works everytime.....dont forget to take it out and warm it back to room temperature before using it

-Jaff-

store at 4C wrapped in damp paper and plastic bag; to use within 2 weeks

-Inmost sun-

to complete the answer, don't leave it set up in the apparatus, with running buffer, overnight. you will allow buffer exchange with the stacking gel (and maybe farther into the running gel). this will affect the stacking and running of the sample (and, hence, the banding pattern).

-mdfenko-

mdfenko on Mon Nov 8 20:20:00 2010 said:


to complete the answer, don't leave it set up in the apparatus, with running buffer, overnight. you will allow buffer exchange with the stacking gel (and maybe farther into the running gel). this will affect the stacking and running of the sample (and, hence, the banding pattern).


Dear an elder,

I came across the problem you mentioned. I kept the gels in the apparatus with running buffer for 3h ahead of leading the samples. When I load the samples, it did not settle in the wells(the loading buffer is not a problem bcos the same has been used by my labmates)as well as I saw some oily matter inside the well. Then I pipted it out and then loaded the samples (the sample settles down perfectly).

Could you advice me whether this is bcos of leaving the gels in the running for 3h or else?

Thanks in advance!

Selvam Ayarpadikannan

-biocrazy-

biocrazy on Sat Nov 20 16:19:59 2010 said:


Dear an elder,

I came across the problem you mentioned. I kept the gels in the apparatus with running buffer for 3h ahead of leading the samples. When I load the samples, it did not settle in the wells(the loading buffer is not a problem bcos the same has been used by my labmates)as well as I saw some oily matter inside the well. Then I pipted it out and then loaded the samples (the sample settles down perfectly).

Could you advice me whether this is bcos of leaving the gels in the running for 3h or else?

Thanks in advance!

Selvam Ayarpadikannan

the "matter" inside the wells could have been polyacrylamide "skins" that get in between the comb and plate when fit is not perfect or dense buffer component(s) that leached out of gel (eg urea, sucrose).

-mdfenko-