Southerns-- Ethidium Bromide

Daniel Meier dmeier at post.its.mcw.edu
Mon Apr 26 09:35:32 EST 1993


Hello,

  After running a Southern (1% agarose, TBE), I added ethidium bromide to
the gel to see the bands, and then transferred the DNA to nylon
(Schleicher and Shuell, Nytran) using alkaline transfer (0.4 M NaOH).
  While the transfer appeared complete, I have a huge amount of ethidium
bromide stain on the nylon.  How can I wash it off?  Will it interfere
with subsequent hybridization?  Is the problem worse with nylon vs.
nitrocellulose?  Finally, Nytran is a positively charged nylon.  Would a
plain nylon membrane be better?
  I will repost all replies.

Thanks in advance,

Dan Meier
dmeier at post.its.mcw.edu



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