Stripping Blots - MSI warning

Robert J. Forster ac562 at FreeNet.Carleton.CA
Fri Dec 8 10:25:32 EST 1995

Dear Probers:

I feel I should make the community aware of a recent problem we have had
with MSI Magnacharge membranes.  After stripping the membranes with 0.4 N
NaOH for 30 mins. (MSI recommends 0.5 M NaOH for 1 hour) we find that the
membranes are well stripped but that YOU CANNOT REPROBE! We had no
problems using this technique in the past and it appears to be related to
a reformulation of the membrane on the part of the manufacturer. 
Stripping with a boiling method (0.1% SDS) or washing in 50% formamide, 6x
SSPE at 65C for 30 mins. strips the oligo probe and allows reprobing.  We
have had no trouble using the NaOH stripping protocol with Hybond membranes.

It took us a while to figure out this problem, checked out our probe
labelling, hyb solutions, crosslinker etc.  and I am rather pissed off
that it was traced down to a change by the manufacturer that was not
passed on to the end users.  Not to mention the loss of valuable samples
and time.  

Cheers, Bob

R.J. Forster					Bob Forster
Centre for Food and Animal Research		ac562 at
Agriculture Canada				tel (613) 759-1725
Ottawa, On K1A 0C6				fax (613) 759-1765

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