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Sample Diffusion Before Applying Current


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#1 mastcells

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Posted 14 August 2009 - 07:40 AM

Hi,
When I add my protein markers to a well, within seconds they begin diffusing down through the gel and somehow into the neighboring wells even before a current has been applied. Any ideas whats going on?

Thanks

#2 cellcounter

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Posted 14 August 2009 - 11:00 AM

View Postmastcells, on Aug 14 2009, 08:40 AM, said:

Hi,
When I add my protein markers to a well, within seconds they begin diffusing down through the gel and somehow into the neighboring wells even before a current has been applied. Any ideas whats going on?

Thanks

This usually suggests that you have some space between your gel and glass plates. Make sure you are using the same thickness spacers and comb when you cast the gel.

#3 mastcells

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Posted 15 August 2009 - 12:03 PM

[/quote]
This usually suggests that you have some space between your gel and glass plates. Make sure you are using the same thickness spacers and comb when you cast the gel.
[/quote]


The strange thing is I'm using precast gels, so I would assume all that should be taken care of, right?

Thanks

#4 THE ROCK

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Posted 15 August 2009 - 09:02 PM

The stacking gel prepared has to be perfect dear B)

#5 cellcounter

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Posted 17 August 2009 - 02:00 PM

View Postmastcells, on Aug 15 2009, 01:03 PM, said:

The strange thing is I'm using precast gels, so I would assume all that should be taken care of, right?

Thanks

I thought I replied to this, but No.

I would suggest that you check how these precasts were stored and handled. i.e.

(1) Expiry date
(2) Drying out
(3) Have gone through some banging while moving them around.





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