in situ with DIG-riboprobe

Frances Hannan Zoology flh at mole.bio.cam.ac.uk
Tue Jan 24 08:36:59 EST 1995


system at cvlab.harvard.edu (CV Laboratory) writes:

>I have been doing in situ hybridization on carotid arteries using
>digoxigenin labelled cDNA.   For some reason, I always get non-specific
>binding with sense probe.

There is a region in the polylinker of the Bluescript vector which hybridizes
with lots of stuff. If your sense probe contains this region it may explain
the non-specific binding to rRNA. The region in question is between the SacI
and SmaI sites and encompasses the SK primer binding site. There is a 
reference to this in NAR or Biotechniques but I don't have it handy. Hope this
helps...

-- 
Frances Hannan                                       
BILMS, Zoology, Downing St, Cambridge, CB2 3EJ, UK   
Phone (0223)336663, FAX (0223)461954                 
flh at mole.bio.cam.ac.uk                               



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