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