I was wondering if anybody has tried (Genius 4) DIG in situ
hybridizations using the Alkaline phosphatase anti-congugate with frozen
sections. Apparently, these sections are levamisole resistant and
methods such as Acid Alcohol, 5mM EDTA/DEPC at 65 degrees, and
increasing Lev. concentration does not decrease this masking background.
I am working with Xenopus tissue from the tadpole stages through
metamorphosis and have not found any literature for doing nonradiactive
in situ hybridizations on xenopus frozen sections. If anyone has any
ideas or leads please pass them on. I am currently looking at other
anticonjugates, as well as, my last resort- radioactive methods.
Thanks in advance,
Chris D'Jamoos
