barley northern normalization

Frank Maiwald maiwald at mpiz-koeln.mpg.de
Tue Dec 15 08:07:29 EST 1998


Dear all,

I am fighting with barley northerns that are meant to show differential
gene expression. One of the problems is to proove equal loading of the
gel or - better - equal blotting of RNA. Many papers that I have
reviewed simply use EtBr- (or acridin orange) staining of the gel / blot
and compare the brightness of rRNA bands. This is ok when you can use
total RNA but fails if polyA RNA must be used. In addition my boss wants
an internal control with a housekeeping gene or some such. The two genes
I tried (alpha-tubulin and S-adenosyl methionine synthetase) did not
give clear signals and SAM is more abundant in roots than in leaves ...

To make a long story short - could anyone give me a hint on a suitable
normalization probe. An accession number would be enough. Then I could
make a PCR probe.

Thanks,

Frank



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