Rad-free kit and genomic southerns

uvsbgsm at Msu.oscs.montana.edu uvsbgsm at Msu.oscs.montana.edu
Fri May 27 14:54:14 EST 1994


I am looking for some advice on using the Rad Free chemiluminescent kit from
S&S for genomic southerns.  I have been trying to get my background down to no
avail.  They advertise it as being very sensitive, but unfortunately my noise
level is so high that it is impossible to tell anything.  If I leave the blot
on the substrate sheet for more than about 30 minutes, things get very black. 
Overnight is completely opaque.  I am doing everything *exactly* as stated in
their directions, and all the tech people can do is scratch their heads and say
its never been like this before.  Even if I take unblotted, unprobed membrane
(neutral nylon provided by them), and detect using their blocking procedure, I
get a black blot in an hour or two.  From this I concluded that I h probably
have a blocking problem.  So, has anyone used this kit and had to adjust the
detection system, and gotten it to work?  Or, am I just hoping to make
something work that won't - as I've been told by some colleagues.  Any help is
appreciated.  Please e-mail me at uvsbgsm at trex.oscs.montana.edu

Thanks!



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