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SDS-PAGE problem: clogged wells, need urgent help

SDS-PAGE problem: clogged wells, need urgent help - SDS-PAGE Gel Electrophoresis Forum

SDS-PAGE problem: clogged wells, need urgent help - SDS-PAGE Forum and Gel Electrophoresis Forum. Discuss the running of agarose gels, sds-page gels, and other gels including sequencing, gradient or tricine.


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Old 08-24-2011, 09:28 PM
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Unhappy SDS-PAGE problem: clogged wells, need urgent help



Hello to the entire molecular biology forum community! I am new here and I hope to find help trough your answers and why not, to help where I can.
So, my question is: why after I remove the comb from the second gel in an SDS-PAGE for plant proteins, and after I rinse the wells neither with distilled water or running buffer, the wells still remain clogged and I can't put the samples into them because the sample does not fall into or fall crooked?
Thank you for your kindness.
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Old 08-27-2011, 05:17 AM
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Default Re: SDS-PAGE problem: clogged wells, need urgent help

hi, sometimes the problem is on the comb itself, which the size does not fit the plate perfectly, leaving some gaps in between the glass plate and comb. Because of these gaps, during casting, thin layer of gel piece will form which cause blockage during sample loading. Another reason could be on the samples, either the sample is too viscous or the density is low, for viscous samples, you can dilute them further while if the sample has low density, you can add some more glycerol into the loading buffer. So 3 things to troubleshoot. Good luck.
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Old 08-28-2011, 01:57 PM
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Default Re: SDS-PAGE problem: clogged wells, need urgent help

Thank you very much for your 3 tips. I think the comb size does not fit the spacers, but the last tip (that with glycerol) is very useful to mee, too. I wish you the best! Have a nice day!
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