DIG In Situs

Asim BZBE at musicb.mcgill.ca
Thu Jan 8 17:59:21 EST 1998


I am using DIG-labeled RNA probes for in situ hybridization to cryostat
sections of brain tissue.  To test the DIG protocol, I used a probe
which had successfully been used for radioactive in situs.  The exact
same hybridization protocol was used with the DIG-labeled probe.

The signal that I got from the DIG probe was fairly low.  While quality
of the tissue may be a major factor, I wanted to know if anything from
the radioactive in situ protocol could be modified when using DIG
probes.  For example, the proteinase K step was fairly short (5 ug/ml
for 5 minutes), but was optimized for the radioactive probe, - would
this affect the ability of the anti-DIG antibody to enter the cells?

Thanks



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