Question: Methods for reprobing nylon filters??

Ed Rybicki ED at micro.uct.ac.za
Tue May 18 08:32:19 EST 1993


> From:          901106c at axe.acadiau.ca (TIM CHIPMAN)
> Subject:       Question: Methods for reprobing nylon filters??
> Date:          18 May 93 12:54:48 GMT

> I have a question: I was curious what methods are available for de-
> colorizing nylon filters (Which have been used with DIG detection), for the
> purpose of re-probing.


...but you don't HAVE to decolourise the membrane...!  That's the nice
thing about the DIG system; you have 2 options of detection, and you can
use both on a single blot without having to decol. in between.  My student
Wendelin Schnippenkoetter has taken a chromogenic DIG blot, boiled it to
get rid of probe, and re-probed with another (different) DIG-labelled
probe, and chemiluminesced the blot: you can then superimpose film and
blot and get a very good idea as to whether the same bands light up or no.
 Works wonderfully - and, of course, there is the option of using
different coloured substrates for AP so as to get multicolour blots....

Forget hot DMF (shudder)!

  ____________________________________________________________________
 | Ed Rybicki, PhD             |       "Lord, won't you buy me        |
 | (ed at micro.uct.ac.za)        |                                      |
 | Dept Microbiology           |         A Mer-ce-des Benz..."        |
 | University of Cape Town     |                                      |
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