[Histonet] Fluorescent in situ hybridization

[Dr. Szigeti Csaba]

Dear Histonetters!

I should optimise Fluorescent labneled DNA oligo probe in FISH
I would like to enhance the specificity of the signal,
My protocol:

2*5 min 2xcSSC wash prehibridization, 0.75% triton, than ON 
hybridization at 30 Grad Celsius in puffer (FISH oligoprobe 25 bp DNA 
fluorescent labeled)
Post hibr wash: 30 min. 10%formamide in 2x ssc 30 Grad celsius+nissl 
counterstain.
Than rinse in 2x ssc and cover.
Result: sometimes weak signal, but very much aspecific dye particles.

Would you have some idea in mind to help me in this protocol. E.g: FA 
fixation at the start is crucial, what you think?

Many thanks in advance to all of you!
Best regards Csaba Szigeti

-- 
DR. SZIGETI CSABA MSC. PHD.

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