Northern Problem

Adrian Molenaar Molenaara at Ruakura.cri.nz
Tue Apr 5 23:00:43 EST 1994


In article <199403252125.NAA05726 at net.bio.net>,
KGEISS%RAVEN at EAGLE.AAMRL.WPAFB.AF.MIL wrote:
> 
> We are currently having problems with our northerns.  It seems that we get a
> lot of nonspecific binding of our probes to both ribosomal RNA bands.  We are
> using cDNA probes for cytochrome P450 2E1.  Any insight into this problem is
> appreciated.  Post or e-mail responses.
> Thanks,
> Geiss...[:)


I suggest you look at the following article, binding to ribosomal sequences
by riboprobes is shown to be due to to CG rich regions found in eg SmaI
sites.

Witkiewicz, H., Bolander M.E., Edwards D.R.  1993.  Improved design of
riboprobes from pBluescript and related vectors for in situ hybridization. 
Biotechniques. 14 458-462.

Another way around the problem may be to increase your hybe temps

Regards

Adrian Molenaar



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