Need help on Northern hyb

Xiaodong Shao shao at umdnj.edu
Thu Jul 23 11:37:01 EST 1992


hi,netters.

I have a problem with Northern hyb using oligos probe. the probe
I used was 55-mers labelled with p-32 ATP. The counting was
200,000,000dpm/ug. Prehyb in 6XSSPE, 0.1%SDS, 10XDenhart's,50ug/ml
DNA. Hyb in 6XSSPE, 1%SDS onernight in65C. Washing 3 times In
6XSSPE, 0.1%SDS 15 mins each. Repeat final wash at 65C. and I got
nothing on the membrance. BTW, the membrance I used was MSI MAGNA
Nylen.

I know there were enough mRNA on the membrance and the mRNA I
needed was moderate expression. 

So what's going on there? any suggestion would be appreciated.

thanks in advance.



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