fli1 transgenics

[wcui at umich.edu]

Hello.

I'm hoping to get a proven protocol for doing double insitu and insitu/IHC
on whole mount embryos.  Here are some questions I have:

1) Can I do double insitu with two DIG-tagged riboprobes, by starting the
second one after finishing the color reaction of the first one?  Or do I
need to get one tagged with FITC?

2) Doing double insitu, what is the best combination of detection
methods--Fast red, NBT/BCIP, other fluorescent tagged antibodies?  And
what's the preferred order of detection if there is any?

3) For insitu/IHC, is there a preferrence in the detection methods?  I
tried to do NBT/BCIP (roche) and HRP/DAB (Vector labs), but the result was
sub-optimal.

That's all.  Thank you.

Wilson Cui
University of Michigan.