RNA stability after electrophoresis

Kyle Legate legatek at hotmail.com
Mon Mar 15 14:03:29 EST 2004


ddao at aecom.yu.edu wrote:
> I am having problems with my Northerns that had worked well in the
> past and was wondering if someone could offer advice. My RNA is not
> transfered onto the nylon memembrane. I think the RNA is degraded
> during the transfer but cannot be sure where the problem is. After
> electrophoresis, I can see undegraded RNA on the gel by UV. But,
> after overnite transfer onto nylon membrane, washed with 2 times with
> 2X SSC, then the membrane is crossed linked by UV. To check for
> transfer, the membrane was visualized by UV. I cannot see the RNA on
> the membrane as I had seen in previous times.  Have tried to probe
> (this probe worked before) the blot and exposed the blot after one 2X
> SSC wash, nothing shows up.
>
Take the rubbery gel and visualize under UV again. Is the RNA still in the
gel?





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