Problems with Southern Blotting

Alicia Subtil aliciasubtil at
Mon May 17 17:27:18 EST 2004

Hi everybody,

I send this message because I am know setting up the coditions to
perform Southern Blotting. Well, the main problem is that I can't
obtain any result with my digested genomic DNA. I have tried the probe
with positive and negative controls that are digested plasmids, but I
don't have any signal with the genomic DNA. I tried it with 15 ug of
genomic DNA but someone told me that he uses 80 ug of DNA!!! This is a
lot of DNA to be ran in a normal agarose gel. What is the amount of
DNA that is necessary to obtain a good signal of a single copy gene in
a Southern Blotting?

Thanks in advance:

Alicia Subtil
Lab. Chromatin and Gene Regulation
Centre de Regulació Genòmica
Barcelona (Spain)

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