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my bands spread as a line trough the gel


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3 replies to this topic

#1 khoshnevis

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Posted 29 August 2004 - 09:10 AM

I loaded my samples in three separate wills, but when I stain the gel, I see some horizental lines in the gel(it seems that the wills leak to each other)

#2 quol

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Posted 29 August 2004 - 11:11 PM

Maybe you overloaded the wells. Try to load less proteins. Of course the amount of protein for good separation depands on many factors as thickness of the gel, number of different proteins... but for example when using BioRad system (1mm gel 9x6 cm, 12 wells) I load not more than 30 mg of cellular extract- works perfectly.
Good luck

#3 joek1

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Posted 30 August 2004 - 06:54 AM

I sometimes see that when the gel is run at a too high voltage. Yet, I agree that you should check to see if you are overloading the wells.

#4 kant0008

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Posted 30 August 2004 - 08:09 PM

Hi,
yes, make sure there's no overloading and so leakage from one well into the next. Also make sure that your spacers and combs are the same thickness- if your combs are too thin you will get gel everywhere, and might be loading your samples into the gel, embedding everywhere, rather than neatly into wells.




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