acridine orange staining

From:Agripina Suarez

Hi Kerry,
I tried on paraffin embedded tissue the acridine orange staining 
protocol for RNA/DNA that you posted in the Histonet a while back. 
I am not getting any brilliant green and orange staining you 
described for results -- only dull orange stain which I suppose could 
be the RNA. You mentioned in your posting that some acridine 
orange dyes work better than others. May I know the source of 
your acridine orange? I got mine from Sigma (Cat # 31,833-7).
Any comments from the Histonetters?

The protocol I followed (posted):
1. Bring paraffin sections to water in usual manner.
2. Stain sections in acridine orange stain for 30 min.
3. Rinse sections briefly in 0.5% acetic acid in 100% alcohol.
4. Rinse sections in additional changes of 100% alcohol.
5. Rinse sections in 2 changes of xylene.
6. Mount sections in Fluoromount.

Acridine orange stain:
Acridine orange --------- 0.05 gram
Dist. water ---------------- 500 mL
Acetic acid --------------- 5 mL

Thanks in advance for your help.


Agripina C. Suarez
Cardiovascular Research Laboratory
Room 292, McDonald Research Wing, St. Paul's Hospital
1081 Burrard St., Vancouver
B.C., Canada V6Z 1Y6
Phone: (604) 806-8659
Fax:   (604) 806-8351 

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