Northerns...again

[Marguerite Evans]

Hi everyone,

Would you believe I am still having problems with Northerns?  I don't 
think my specific problem is related to the method now, but to my 
probes.  Has anyone experienced (with either a Northern or a Southern), 
upon re-probing the membrane, that they get their probe binding all over 
the place in "splotches" if you like.  It is like it is binding to 
something else on the filter - could this be dried up SDS that sticks 
well to the filter?  (I strip membranes by boiling 0.5% SDS, pouring it 
onto the membrane and letting it come to RT.  I then blot the excess from 
the membrane using 3MM and re-probe.  I haven't seen this before and it 
is just ANOTHER item to add to my long list of "Northern problems".  My 
probes bound beautifully (if you could say that) to the spots...in that I 
cannot get it off!!  

Still I repeat the mantra passed onto me by Mike Leibowitz..."there's no 
such thing as a trivial Northern blot....there's no such 
thing...."

Thanks for listening...
Maggie.

Marguerite Evans School of Biochemistry and Molecular Genetics
UNSW  Sydney  2052
NSW
Australia

voice: (+61)(2)385 2030
fax:   (+61)(2)313 6271
email: p2158740 at acsusun.acsu.unsw.edu.au