RNase protection assay

Patrick F.H. Lai the Graduate Student pfhlai at LOOKSMART.COM
Wed Oct 14 00:36:33 EST 1998

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Does anyone know what a RNase protection assay is and what is its advantage
over filter hybridization in quantitating gene transcription?



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---- End Original Message ----

Dear Dan and Netters,

This is how I understand what RPA is, correct me if I am wrong:

RPA is a simple way of detecting and quantitating RNA instead of doing a Northern blot.  Basically, a anti-sense probe (typically a few hundred bases long) is mixed in with your sample of RNA extract.  Probe binds to your RNA of interest and is protected when you throw in ssRNA-specific RNases.
Run a gel and transfer.  If your probe is radiolabelled or fluorescent-tagged, you will see a band on your blot.  You design your probes, so you know what size of the band to expect.

Advantage ?  One can detect several RNA at the same time.
Simply use probes of different sizes that one can resolve in a gel.

Check out www.ambiom.com
I heard that their newest kit even allows up to 6 RPA in one shot!

BTW, has anyone actually took up Dr. Eric Lader's offer to try out their RNAlater! solution ?  
How is it ?

Hope this is useful info.   :-)

Patrick F.H. Lai  < PFHLai at looksmart.com >
Graduate Student
University of Toronto
Toronto, Ontario, Canada

P.S.  I am merely a GradStudent.
One should take a HUGE grain of salt with whatever I have typed above.
Good Luck.   :-)


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