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Position: Discussion forums > Bioscience biotechniques discussion forums > Immunohistochemistry In Situ Hybridization forum
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RNA in situ hybridization
Posted by: Arielle (IP Hidden, Unregistered user, )
Date: April 11, 2005 05:31AM
Hi!
I perform RNA in situ hybridization on paraffin-sections. I use the Roche Nucleic Detection Kit with DIG Labelled probes, AP-conjugated Ab and NBT/BCIP. My problems occur when i stain the sections. After staining the tissue turns brownish but after stopping the reaction and waiting a few days the tissue turns blue.. This wouldnīt be that much of a problem if it wouldnīt keep on turning darker and darker and after a few days itīs so dark that itīs useless as no distinct cells can be seen anymore. This problem occurs with antisense and sense probe, so even if the sense probe is negative at first it turns positive after some days.. Has anyone had this problem before? I really need to store my slices for some time as I have to compare them at the end of the project. Taking picture at once wonīt help me much.. Thank you for your help Arielle
Re: RNA in situ hybridization
Posted by: affaro (IP Hidden, New member, 2)
Date: July 19, 2006 07:47AM
Hello
I have a similar problem to yours. Did you found a solution yet? Thanks.
Re: RNA in situ hybridization
Posted by: marina (IP Hidden, New member, 1)
Date: October 9, 2006 04:23AM
Hi!
I have a general question. I perfofm in situ hybridization, using DIG-labeled probes. When I move to the revelation step, I get a different colour of staining every time. For example, some times my staining is blue, or other times brown, and the last time it was yellowish! Does anyone know why this happens and how I can avoid it? Thanks.
Re: RNA in situ hybridization
Posted by: shijianlixj2006 (IP Hidden, New member, 2)
Date: January 21, 2008 08:18PM
Hi!
I perform whole mount in situ hybridization on Xenopus embryos, but I always meet many macula after staining.Does anyone know why it happens and how I can avoid it? Thank you
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