Destaining NBT-BCIP -northern-blot

Michael Witty mw132 at mole.bio.cam.ac.uk
Tue Jan 23 07:47:27 EST 2001


Dear Francoise, I did not know that you could remove the indole coloured
precipitate from NBT-BCIP stained membranes.  How did you find out about
this?  It sounds good for my NBT-BCIP stained western blots!
Regards, Mike.

On 23 Jan 2001 francoise.martz at metla.fi wrote:

> Non-radioactive northern-blot, colorimetric NBT-BCIP detection:
> To reprobe a northern membrane (nylon N+) using the DIG and the 
> colorimetric system, I am removing the precipitate by incubation in DMF, 
> 50oC, but there are 2 problems:
> - strong staining is not removed,
> - I have the feeling that RNA are also removed (they are fixed by backing 
> 80oC, 2h).
> So is there any other possible treatment to remove the color precipitate, 
> that keeps the RNA on the membrane...?
> Thanks for your help.
> Francoise
> 
> ___________________________________
> Françoise Martz
> The Finnish Forest Research Institute
> Rovaniemi Research Station
> P.O. Box 16
> 96301 Rovaniemi, Finland
> Tel: +358 16 336 4215 - Fax: +358 16 336 4640
> 
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