purification of radioactive probe
(by k.schinkNo from SpamGmail.com)
Tue Aug 28 07:13:50 EST 2007
Simone Marker schrieb:
> 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,
We purify our radioactive probes using small gel filtration spin columns
(I think it's called MobiSpin) - you get rid of most unincooperated
radioactivity, which is retained in the column, while your labelled
probe is in the flow-through. It makes it much nicer to work with the
probe because it is not as radiactive as the whole reaction mixture..
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