In situ hybridization probes

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From:Mike Bromley <> (by way of Marvin Hanna)
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Dear All,
I have been staining formalin fixed paraffin embedded tissue sections
for Epstein-Barr encoded small RNA's (EBER's) using the method of
Helene Breitschopf from the Boehringer manual: Non-radioactive in situ
hybridization  application manual 2nd edition.(published also in Acta
Neurapathol (1992) 84:581-587).

The method uses digoxigenin labelled RNA probes, the anti-sense is
giving good strong staining in the right cells, unfortunately the
sense probe is also staining the same cells albeit not as strongly.

In the Boehringer manual p137, it states that hybridization with sense
probes sometimes gives unexpected hybridization signals.

Has anyone got any ideas about why the sense probe should apparently
be specifically targetting the right cells?

Mike Bromley
Histology Department
Paterson Institute
Wilmslow Road
M20 9BX

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