PROTOCOL
1. Incubate chicken eggs for 2 days.
2. Remove eggs from incubator. Aseptically window eggs(do
not remove membrane) and inject control and experimental
eggs with 1uL of solution into yolk and away from
embryo.
a. Control egg solution: 1mL
of Howard
Ringer's solution, 100uL
DMSO (5
mM)
b.
Experimental egg solution: 1mL of Howard Ringer's Solution,
100 uL
SU5402* (5 mM).
*handle
SU5402 with extreme caution!
3. After windowing and injecting the control and
experimental solutions, place in 37ºC incubator for 1
or 2 day(s).
4. Open eggs and remove embryo aseptically. Clean embryos in
Howard Ringer's Solution and capture images of experimental
and control embryos.
5. Preserve in 4% Paraformaldehyde.
6. Process embros with DIG-labled
shhriboprobe
formin
situhybridization.
7. Capture images ofin
situhybridized
experimental and control embryos.
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