rRNA Hybridisation in NB

Anke Wichert anke.wichert at charite.de
Wed Oct 6 10:38:45 EST 1999


Hi all,

I have never seen this problem before, although many of my colleagues
had (and I always thought them just messy or so), anyway, now I faced
this: after completely and finely blotting a close to perfect RNA
agarose gel, I hybridised it with a denatured DNA probe (Amersham
megaprime kit). The same had worked well in my hands for dozens of
times. However, the autoradiography showed the rRNA bands like in the
gel, but not my desired mRNA. As I said, the total RNA as well as the
probe and kit  had worked fine before. I tested the probe's DNA on an
agarose gel, and it was as sharp as a knife. I am repeating the NB, but
am strongly interested in the problem as it did occur with others
before.

Thank you most cordially!
Anke




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