Help spotty northern problem help

punk at leleand.stanford.edu punk at leleand.stanford.edu
Tue Aug 5 13:22:09 EST 1997


Hi, I am doing a lot of northerns on cellular RNA and having a problem
with spots on my autorads. These spots are small and round. The
hybridization signal is good, no crosshyb to rRNA and the general
background is low except for the spots. I am using  zeta-probe membrane
from biorad and have changed lots which didn't help

- I am using the .5M NaPO4  7%SDS  hyb buffer. It used to work well no
spots and this is what is recommended for the zetaprobe.  I have switch
the solutions and filtered them but still the spots are present.

- I am using random primed probes and a hybridization oven. The probe is
cleaned up using qiagen spin column 

If any body has any suggestions on what might be causing these spots
please post to this group I check it regulary Thanks alot gregg



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