non-radioactive Northern blotting

Neil Saunders saunders at bio.vu.nl
Tue Oct 27 08:29:23 EST 1998


I've used random-primed DNA probes with some success in Northern
blotting of Gram-negative bacterial mRNA transcripts, but only where the
transcript of interest was at a relatively high level (enough to show up
at 10 microgram total loading or so).  I've recently switched to
DIG-labelled riboprobes and found that they're much more sensitive.

I also use the detection protocol of:

Reduction of background problems in nonradioactive northern and Southern
blot analyses enables higher sensitivity than 32P-based hybridizations.
Engler-Blum G, Meier M, Frank J, Müller GA
Anal Biochem 1993 May 1 210:2 235-44

which works well for me.

Neil Saunders

Vrije Universiteit, Amsterdam





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