7 replies to this topic

[Aris]

I am preparing my own gels with a new recipe that is acctually very good. Still lately I have noticed that in the wells after i remove the coumd, there are some fibres that make tha loading very difficult.
Has anyone any experience with this odd thing? Is there a way to avoid them??

Thx a lot

[bob1]

solid fibers or semi-liquid? My guess is un-polymerised acrylamide. The resolution is to rinse the wells out several times with distilled water after taking the comb out. You need to squirt the water in reasonably hard to get rid of the acrylamide.

[T C]

Hey,

I faced a similar problem once and the mistake was that I was using 0.5 mm combs and the cast was meant for 0.75 mm so the comb used to hang in between acrylamide and when the combs were pulled out, it had these threads and thus problem in loading.



Best,
TC

[Aris]

bob1, on Mar 26 2009, 04:56 PM, said:

solid fibers or semi-liquid? My guess is un-polymerised acrylamide. The resolution is to rinse the wells out several times with distilled water after taking the comb out. You need to squirt the water in reasonably hard to get rid of the acrylamide.

I am using the right comb for the Gel, its 1,00 mm, i have already washed the gel couple of times but the problem remains. Its as you say semipolymerised acrylamide. The one think i dont know is whether you wash them before you assemble tha apparatus and put running buffer or afterwards. Or doesnt it make any difference....

[Andy Kralik]

Hi.. have you found any resolution of this problem?

Maybe it could help me - I have the same problem with the fibres in the well. The loading of the samples is then very complicated. I thought that the problem was in buffers, I tried to prepare new, but the fibres remain.

I does not depend on that if I use 1mm or 1,5mm spacers and combs.

Edited by Andy Kralik, 04 July 2009 - 06:00 AM.

[bob1]

Andy Kralik, on Jul 4 2009, 06:59 AM, said:

Hi.. have you found any resolution of this problem?

Maybe it could help me - I have the same problem with the fibres in the well. The loading of the samples is then very complicated. I thought that the problem was in buffers, I tried to prepare new, but the fibres remain.

I does not depend on that if I use 1mm or 1,5mm spacers and combs.

Wash your wells out with water, squirt it into the wells and then tip it out, so that the wells empty. This will involve shaking the assembly to get rid of the residual water.

[Moses]

i have a simlar experience too. the unmatching comb for cast is surely the problem.

T C, on Mar 27 2009, 11:31 AM, said:

Hey,

I faced a similar problem once and the mistake was that I was using 0.5 mm combs and the cast was meant for 0.75 mm so the comb used to hang in between acrylamide and when the combs were pulled out, it had these threads and thus problem in loading.



Best,
TC

[mdfenko]

this problem can also occur because of warped plates.

we physically scrape out the well with a thin, flat spatula. another lab aspirates the "skins" from the well.