postmast at iihema.fmed.uba.ar postmast at iihema.fmed.uba.ar
Fri Jun 21 22:31:41 EST 1996

songj at aa.wl.com (Jackie Song) wrote:
>Hi!  I am trying to incorporate 32P-CTP into my PCR probe with no luck.  
>I have been using my regular PCR protocol except with hot CTP and when I 
>probe my Northern I get no results.  Does anyone have a proven method for 
>this?  How hot should be probe be?  Is there any easy way for me to 
>ensure the hot CTP is being incorporated into my product?	
 2 microCi of 32P-CTP should be enough to get a strong signal.
 To check whether 32PCTP is being incorporated or not you could run the
 product in an agarose gel, expose the gel to a film and see if you get a
 fragment the desired size in the autoradiography.
 Are you sure your probe matches correctly the RNA your searching in your 
 Are you sure you're doing the northern correctly (right hybridization 
 temperature, etc.)?
 Hope this helps,
 Facundo Garcia Bournissen,
 postmast at iihema.fmed.uba.ar

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