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making gel in advance


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7 replies to this topic

#1 SF_HK

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Posted 06 November 2010 - 12:21 PM

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?

#2 BioMiha

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Posted 06 November 2010 - 01:48 PM

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.

#3 ksturm

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Posted 06 November 2010 - 03:33 PM

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

#4 Jaff

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Posted 08 November 2010 - 06:30 AM

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

#5 Inmost sun

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Posted 08 November 2010 - 10:22 AM

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

#6 mdfenko

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Posted 08 November 2010 - 12:20 PM

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).
talent does what it can
genius does what it must
i do what i get paid to do

#7 biocrazy

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Posted 20 November 2010 - 08:19 AM

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

#8 mdfenko

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Posted 22 November 2010 - 12:29 PM

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).
talent does what it can
genius does what it must
i do what i get paid to do




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