nonradioactive rna probes

Tracy Aquilla aquilla at salus.med.uvm.edu
Mon Jan 23 15:47:56 EST 1995


In Article <3fpl0e$mgn at netnews.upenn.edu>, aaron at netnews.upenn.edu (Aaron
Pawlyk) wrote:
>Fiberman (william at neuro.usc.edu) wrote:
>
>: I want to run an RNAase protection assay as opposed to a Northern so
>: I can avoid transfer.  But is there a way to detect the bands using
>: nonradioactive method?  I need to visualize the bands in the gel
>: as opposed to on a membrane.
   ^^^^^^^^^^^^^^^^^^^^^^^^^^^
>
>: -fm
>
>You should look into the non-radioactive RNA probe systems commercially
>available.  I believe Promega, Invitrogen and Ambion all have systems available
>for non-radioactive probes.  You can use them as you would in a normal RNase
>protection assay.
>
>The only problem is the the sensitivity of these probes are much lower than you
>maximally achieve with a 32P.  So if you need a highly sensitive RNase
>protection assay it won't work.  
>
>                        -- Aaron
>

Well, I don't think that's the only problem. The non-radio techniques I'm
aware of require blotting the gels and staining with chromogenic substrates
or using chemiluminescense. (Did you read the question?) This would defeat
(much of) the purpose of doing RNAse protection assays.
    Tracy



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