Michael L. Sullivan
mlsulliv at facstaff.wisc.edu
Mon Jul 16 09:47:28 EST 2001
>> The problem with this strategy is that your EtBr will migrate in the
>> opposite direction of your DNA during electrophoresis. This will make
>> that the band with a lower molecular weigth will be less visible.
>> Soaking the gel in a 1ug/ml EtBr solution for 10 min eradicates that
>We find also that adding EtBr directly to the running buffer as well as
>the gel eliminates this problem. No soaking, destaining, or other annoying
>step is necessary. Just add EtBr to the same concentration as in your gel.
>Background is very minimal in our experience.
Most of the time I think the EtBr can be omitted from the running buffer
since most people are so anxious to see their results gels seem to rarely
get run more than half way.
Even if a gel is run all the way without EtBr in the buffer, although the
bands are much lighter, the background is extrememly low, so I've found
that it's still easy to detect bands.
More information about the Methods