Differential Display Screening

Brian btreacy at afns.ualberta.ca
Thu Aug 20 12:41:19 EST 1998


  Hi Everyone!


  I have recently cut out over 30 unique bands using GenHunters RNA Image
kit. They have all been cloned in GenHunters pCR-TRap. I have tried both
recommended approaches (i.e. dot blot of PCR products and colony
hybridization) to no avail. My autorad shows these "halos" surrounding
either the colony streak or PRC product loaded!! I have taken precaution in
denaturing both sample and probe followed by quick cool. Has anyone ever
seen or heard of this? My probes were 45 000 CPM after G-50 sephadex and
were added to my hyb buffer at one million counts per ml. I'm thinking
northern blots and selecting a few clones from my collection of 30 as
probes. I'm sortta stumped here!

  Thanks crew,

Brian

Brian K. Treacy, Ph.D
Agriculture, Food and Nutritional Sciences
University of Alberta
Edmonton, Canada





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