EtBr in agarose gels...

Kirsty Kirsty
Tue Feb 28 04:10:35 EST 1995


In article <Pine.A32.3.91.950222182822.74692A-100000 at rs5.tcs.> Brick Ayola <abrick at mailhost.tcs.tulane.edu> writes:
>
>Regarding EtBr staining of nucleic acids:
>Can one simply add EtBr to the loading dye (as with some RNA gel 
>protocols)? Or will this significantly disturb the migration of DNA 
>super-coils and such?
>
>Brick Ayola, Ph.D.
>Tulane School of Medicine

Brick,

You can either add EtBr to the agarose before setting or stain the gel after
electrophoresis, either way will not alter the DNA migration in conventional
electrophoresis. However, with pulsed field gel electrophoresis you must stain
the gel with EtBr afterwards rather than add it to the gel. DNA migration will
be significantly altered if EtBr is added at any stage prior to electrophoresis.
For best results, stain with EtBr for ~20 min then destain in distilled water for 
~20 min.

Kirsty Townsend
Molecular Oncology
QEII Medical Centre
Nedlands  6009  WA
AUSTRALIA
ktownsen at eosin.path.uwa.edu.au



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