fluorescent RNA stain for microscopy

Frank O. Fackelmayer fof1 at chclu.chemie.uni-konstanz.de
Mon Nov 25 03:58:16 EST 1996


In article <199611221539.JAA00623 at borcim.wustl.edu>,
brett at BORCIM.WUSTL.EDU (brett) wrote:

> >Hi everyone!
> >IYm looking for a fluorescent stain that is specific for RNA (no DNA
> >staining!) and can be used in fluorescence microsopy of nuclei. What I
> >would like to have is something like Hoechst 33258 or DAPI or EtBr, but
> >for RNA. As I donYt want to visualize a specific RNA (or poly A-RNA), in
> >situ hybridization cannot be used. Rather, all types of RNA should be
> >stained. 
> >
> >I know of some chemicals that light up DNA and RNA, but with different
> >colors (e.g. acridine orange and derivatives). However, this is NOT what I
> >want to use unless absolutely necessary.
> >
> >Any hints, references, and suppliers are appreciated!
> >
> >You can reach me directly via e-mail at:
> >fof1 at chclu.chemie.uni-konstanz.de
> >
> >-- 
> >Dr. Frank O. Fackelmayer
> >Division of Biology
> >University of Konstanz
> >D-78434 Konstanz
> >Germany
> 
> You could load your cells with BrUTP prior to fixing, and detecting with
> anti-BrUMP.
> 
> 
> Brett Lindenbach
>     
> Program in Immunology                              
> Washington University - St Louis                  
> brett at borcim.wustl.edu                             


Thanks for your suggestion. I think it would be good for all purposes
except mine. WHY? My problem is that I don´t have cells, but assemble
nuclei from scratch by using Xenopus laevis egg extracts (these extracts
can build up nuclei around any DNA you add, so they are a perfect model to
study nuclear assembly). I want to investigate whether the RNA in the
extract (which is VERY abundant), is incorporated into the nuclei.
Labelling RNA with BrUTP cannot be used, as these extracts do not
transcribe (Remember its an embryonic system that has enough maternal RNA
for several cell cycles!). 

Any other suggestions?
Frank

-- 
Dr. Frank O. Fackelmayer
Division of Biology
University of Konstanz
D-78434 Konstanz
Germany



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