RNA isolation

susanne srohrerNOsrSPAM at immv.unizh.ch.invalid
Fri Nov 19 10:12:12 EST 1999


> If you want to transfer large bands you should treat gel
> (especialy thick
> ones) with 0.05N NaOH (1ml 10N +200ml H2O) for 15 minutes exactly
> prior to a
> few water rinses, pyrex exchange, 20XSSC 45mins with a change in
> the middle.
> If your blotting was good, a picture of the "pancake" will show
> nearly no
> ribosomal RNA.

Are you sure Alkali treatment is a good idea? I have switched to
neutral blotting because I felt my larger transcripts were being
degraded.

After running the gel, I rinse 2x15min in 20xSSC.

Susanne Rohrer, Uni Zurich


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