Protocol Online logo
Top : New Forum Archives (2009-): : Molecular Biology

Nonidet P-40 (NP-40) in washing buffer after hybridisation - (May/04/2012 )


the following FISH protocol after probe hybridisation is given:

Inkubate FISH samples 2 min in washing buffer 1 (0.4x SSC / 0.3% NP-40) at 72°C. Afterwards incubate for 1 min in washing buffer 2 (2xSSC/0.1% NP-40) at room temperature.

My questions are:
What is the function of nonidet P-40 in this washing buffer?
Why do you use SSC and no other buffer for example PBS?

Thank you very much in advance. Until now I haven't been able to find anybody you could help me.



Nonidet P-40 is NOT NP-40. This is a very common mistake in all labs I have ever seen.



SSC is a stringent buffer - check out the pH and other conditions.