[Histonet] ISH probe labelling

From:"Mikael Niku"

Dear Histonetters,

I'm about to start doing some tissue in situ hybridization for a
not-so-abundant mRNA target.
I'm uncertain about the probe labelling method I should use. I need to
use an oligonucleotide probe. I have previously used an oligo probe for
a highly amplified genomic target, and cRNA probes for mRNA targets. But
I don't have any experience in this particular combination.

So what do you think:
Will a probe with a single DIG label be sensitive enough?
If not, is it feasible to order a doubly or triply DIG labeled probe, or
should I try tailing instead?

I will be using either the typical anti-DIG alk phos + NBT/BCIP
detection system, or tyramide amplification, if necessary.

With best regards,
Mikael Niku

+  Mikael Niku		 
+  University of Helsinki, Dept. Basic Veterinary Sciences
+  URL: www.helsinki.fi/~mniku/

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