RNA stability after electrophoresis

ddao at aecom.yu.edu ddao at aecom.yu.edu
Sun Mar 14 23:44:10 EST 2004

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.

What do you think could be happening? Could it be related to formaldehyde in the gel? 

Thank you for your responses.



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