[Pombelist] FISH protocol

[Jikai Wen]

Hi Everybody,

I am going to do FISH in S.pombe to check my RNA localization by four 50 nt specific probes, which have C6 amino-modification on dT and could be labelled by Cy3. I get the yeast FISH protocol from my collegue that works excellent in budding. I am wondering that what's the difference for the cell spheroplasting between fission yeast and budding yeast, simply in budding, they always use lyticase and perform it in ~10 to 15 minutes after fixation but in Nurse-lab protocol, they use zymolyase and novozyme with 90 min incubation.

Does somebody have the FISH protocol in pombe?

Thank you very much!
Cheers,

Jikai