soutern problems

Bike&Linux jaudall at students.wisc.edu.nospam
Wed Feb 25 00:24:22 EST 1998


I'm woking in a verteran Southorn blotting/RFLP lab but recently we have
had a wierd problem come up that has us stuck.  We're getting portions
of our blots very hot (black).  If there was more than one blot in the
hyb bottle, these portions can be put together to represent the blot
pieces (i.e. strips on the autorads) which were in contact with the
glass.  I'm hoping this information enough my trigger somone's memory
who has had this problem before.

We're using primer extention (random hexamers) and subsequent
centrifugation through a sephadex spin column to remove un incorportated
nucleotides.  I realize there are soooo mant different parameters to
RFLPs and a complete layout of our methodology might be helpful - but I
hope someone has ran across this situation before.

Anyone?

Josh Udall




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