In article <00970394.67123A40.12574 at happy.uccs.edu>, slblowe at HAPPY.UCCS.EDU writes:
> Dear networkers,
> Is anyone out there screening libraries? What do you use to
> denature and neutralize your filters? We place them on saturated
> Whatmann 3mm paper, but would like to find a replacement because
> of the cost of 3mm. Does anyone know why 3mm is used? Is it
> because it has a neutral pH? Any other, less expensive, suggestions
> would be welcome.
> Sandy BerryLowe at Univ. Colorado
> email responses to slblowe at uccs.edu.>> Thanks
We just put our filters directly onto the buffers.
We use petridishes of the size of filters fill them with buffer and put the
filters upside down onto the liquid surface (try to avoid bubbles underneath)
for 5 min each (denat. buffer: 500 mM NaOH, 1.5 M NaCl; neutr. buffer: 1 M
Tris-HCl pH 7.0, 1.5 M NaCl), then let them dry on filterpaper (upside up :-)
and crosslink. Thats all - and you can reuse the petridishes. We never had any
problems with this procedure - hope its usefull for you.
Max-Planck-Institut for Molecular Genetics