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In situ hybridization enigma - (Sep/03/2010 )

My in situ hybridization is a nightmare!!!!!
I perform it using a protocol that has worked very well for the first and second experiments. Then for next experiments the signal is very low and unspecific. I have not change the RNA probe and the tissue sample. Do anyone know which could be the weak link ?
Gabriella

-gabriella-

gabriella on Fri Sep 3 11:06:24 2010 said:


My in situ hybridization is a nightmare!!!!!
I perform it using a protocol that has worked very well for the first and second experiments. Then for next experiments the signal is very low and unspecific. I have not change the RNA probe and the tissue sample. Do anyone know which could be the weak link ?
Gabriella


Make a new probe. Check that your other reagents are RNAse-free, best way is to make all of them new.

Cheers,

rsm

-Rsm-