Southern blots

vivanova at my-deja.com vivanova at my-deja.com
Tue Nov 30 13:31:22 EST 1999


HI, all!

Here I have a question to all the Southern experts:

What's the best way to do a Southern transfer? Maniatis uses a HCl
followed by 0.5 N NaOH to depurinate and nick the genomic DNA, then
transfers in SSC. Has anyone of you got a protocol that uses NaOH as the
transfer buffer? Any useful suggestions and comments would be greatly
appreciated!Previous experience has given somewhat fuzzy bands om the
film. Could that be improved in some way? Is the HCl/NaOH contributing
to the fuzziness?

Thanks a lot!

Vesselina


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