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Cell lysate stuck in gel well


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#1 Jon Law

Jon Law

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Posted 20 October 2010 - 02:55 AM

Hi, I'm new enough to running gels so this is probably an obvious problem with an obvious answer but here goes anyway.

I'm analysing amplification from PCR on a gel and it works fine when I'm performing RT-PCR from isolated RNA samples. Distinct bands are visible on the gel. However when I perform RT-PCR directly from unpurified cell lysates, the sample seems to remain trapped in the wells on the gel. I presume that this is because of proteins, gDNA and general junk that would be present in the cell lysate and is inhibiting migration. Is this the case and if so, is it a common problem? Is there any literature available regarding this as it would be helpful when I am writing about it?

Thanks in advance!




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