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southern - loading DNA (Apr/18/2006 )

hi
to run DNA for a transfert, do I need to to add ethiBromid to the gel and do I need to add loading buffer to my DNA sample.
does it onterfer with the southern
thanks

-ulujm-

hi
EtBr should be loaded with the sample for visualization. It does not interfers with southern (same as in northern assays).

-fred_33-

as far as loading buffer...depends on your detection method.
sometimes with chemi detection, I get a little bit of smearing across the gel where the loading buffer was...so I usually just dilute it 1:10 and add a very small amount (you still need your sample to sink into the well, yeah?)

by the way, the 'smear' I sometimes see from where the blue juice transfers to the membrane, is clearly distinguished from a good tight band...but looks tacky if you might ever want to publish the results

-aimikins-