Novocrania in situ en, wnt1, wnt5


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Started an in situ with Novocrania! I picked 3 specimens of each stage making 12 embryos in total. The stages are: radial, bilateral gastrula, early larva and larva.

Nano en Nano wnt1 Nano wnt5

All 12 embryos per well survived the first day.

25/09/13

Since engrailed probe had low concentration I diluted what I had (13 ng/µL) into 200 µL of hybe (resulting in 0.5 ng/µL) and added anyway. I’m synthesizing more so that I can add tomorrow.

26/09/13

Changed engrailed hybe with nice new probe. However, embryos are really transparent. Larval morphology also does not look normal. I think something is wrong.

27/09/13

Engrailed well has not any normal looking embryos anymore. They are all falling apart. In the other 2 wells there are still 4-5 embryos that look ok, but fragile.

I continued with the washes anyway and incubated with the antibody overnight at 4 °C.