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.
EGYETEMI ADJUNKTUS
Szegedi Tudományegyetem Általános Orvostudományi Kar
Anatómiai Szövet-és Fejlődéstani Intézet
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