nonradioactive northerns

Jodi Lindsay jodi at arbo.microbiol.uwa.oz.au
Fri Nov 20 03:00:16 EST 1992


In article <1992Nov17.142120.26717 at gserv1.dl.ac.uk> SHANELT at BIOLAN.UNI-KOELN.DE (Sabine Hanelt) writes:
>From: SHANELT at BIOLAN.UNI-KOELN.DE (Sabine Hanelt)
>Subject: nonradioactive northerns
>Date: 17 Nov 92 14:15:17 GMT



>Hi netters,
>my problem is how to get a nonradioactive northern blot.
>I tried already Digoxigenin DNA and RNA probes (Boeringer System)
>and also biotinylated DNA probes (USB Gene Images). None of these
>systems seem to be helpfull, maybe I make a general mistake.
>Has anyone experiences with nonradioactive northern blots that are not
>so frustrating like mine?

>Thanks for your help
>Sabine



Just a suggestion - have you heard of the Amersham Enhanced 
Chemiluminescence Gene Detection System??  (yes...another kit!!)

It involves labelling probe with horseradish peroxidase using 
glutaraldehyde.  Detection occurs by washing the hybed membrane with luminol 
and peroxidase (and enhancers) which causes a chemical reaction that gives 
off light, which is captured on x-ray film.

Although the standard procedure uses DNA probes it is being successfully 
used with ribosomal RNA in our lab.  The method is fast and sensitive, 
although the detection kits are prone to going off, and are rather 
expensive.  We have not been able to figure out what the "enhancers" are 
so as to make our own.

Regards

Jodi Lindsay
Department of Microbiology



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