purification of radioactive probe

Jayakumar, R via methods%40net.bio.net (by R.Jayakumar from roswellpark.org)
Mon Aug 27 11:48:51 EST 2007


Helps reduce non-specific signals.  If I remember correctly, the random
primers sometimes generate much smaller labelled fragments (that is whey
they are random) which can give you a lot of non-specific bands in your
northern blot.  I used to use some sort of spin columns which did not
take more than 10 minutes of my time.  It helps a lot.  
   Best of luck
Jay


-----Original Message-----
From: methods-bounces from oat.bio.indiana.edu
[mailto:methods-bounces from oat.bio.indiana.edu] On Behalf Of Simone Marker
Sent: Monday, August 27, 2007 9:06 AM
To: methods from magpie.bio.indiana.edu
Subject: purification of radioactive probe

Hi,

does anyone know if it is really necessary to purify a radioactive DNA
probe after labelling it by random priming? I want to use it on a
northern blot. 
So far, I worked with DIG-labelled probes, that do not need to be
purified after labelling. They are used directly in the hybridisation
solution. Is it different with radioactive probes? Do you think that
precipitation by salt and alcohol is sufficient for purification?

Thank you very much,
Simone 


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