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purification of radioactive probe

purification of radioactive probe - Protocols and Methods Forum

purification of radioactive probe - Post Any Protocol, Method, Technique, Procedure or Tips / Troubleshooting for any Molecular Biology Technique.


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Old 08-27-2007, 04:48 PM
Jayakumar, R
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Default purification of radioactive probe



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: [Only registered users see links. ]
[mailto:[Only registered users see links. ].indiana.edu] On Behalf Of Simone Marker
Sent: Monday, August 27, 2007 9:06 AM
To: [Only registered users see links. ]
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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