Protocol Online logo
Top : Forum Archives: : Molecular Biology

RNA in situ hybridization - Whole mount RNA in situ hybridization on vertebrate embryos (Oct/04/2005 )

Hi everyone,
New to this forum.
Having trouble in getting good signal during non radioactive RNA in situ hybridization on whole embryos (mouse).
Following the standard Wilkinson's protocol,but everytime a diffuse backgroung comes all over the specimen being used.
Please suggest me how to improve signal intensity and how to get rid off background.



Please look at this wabsite, it might provide some clue.

Best regards


Thanx for the suggestion Hadrian,
Actually i have already gone through this site, infact i've read most of the literatures available,but the problem is......earlier i used to get very gud results in in situ, but from the past few days, i'm trying n trying, and not getting gud signal.
've changed all the reagents, all stocks n everything, gone through each n every step to look for shortcomings, but i'm unable to trace out what mistake i'm doing...................
the bottom line is that, the complete tissue gets a diffused stain as soon as i add the substrate, there is no difference in the signal and background.

i'm very anxious to hear any sort of suggestion regarding this.
Can anyone tell me what cud be the possible reason???????