In situ Novocrania and Terebratalia
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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.