In situ Novocrania and Terebratalia 

In situ using Nano longer probes to try to get a better signal to ratio. Also fgf8 probe, important for the brain boundary. I also need better hedgehog pictures, so I’m repeating the in situ with a new probe in Ttra and also trying to get a good early gastrula, bilateral gastrula e late larvae of engrailed.

Nano wnt5 Nano smo2 Nano fgf8 Nano ptc1
Ttra hh Ttra en late blastula / early gastrula Ttra en bilateral gastrula / late larva

10 min ProtK for all. Incubated at 65 °C at 2000.

31/01/2015

Diluted probes to 1 ng/µL and added at 0800.

02/02/2015

First day of washes. Added anti-dig-ap at 1600.

03/02/2015

Washed off antibody with 3x 5min followed by 4x 15 min and 5x 30 min. Developed with nbt/bcip at 1600. All wells showed good signal/noise ratio. The worst is wnt5, but it is still better than the other in situ. I exchanged the AP for all wells at 1930 and stopped all except hedgehog and en bilateral (exchanged AP).

04/02/2015

Exchanged the AP for the two Terebratalia wells, I’ll let them develop longer.

Stopped at 1500 and did ethanol washes. Let overnight in 70% glycerol.

05/02/2015

Mounting and pictures.