Terebratalia and Novocrania in situ 


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Began in situ 10 min protk (half-concentrated for Nano) with:

Ttra wnt2 Ttra wnta  
Nano wnt8 Nano wnt11 Nano pax2/5/8

07/12/13

Added probes at 13:00.

09/12/13

First day of washes, added anti-dig-ap around 18h.

10/12/13

Washing off antibody, starting with 2x 15min PBT and after 1h 3x 15min PBT and normal PTw washes.

Started developing at 16h. Some signal started to appear at Wnt11 ~18h30.

11/12/13

Arrived 9h30 and the wells were purple! Did an extra wash with AP substrate and let at RT. Exchanged AP 4 times and stopped the reaction at 19h with 3x AP without MgCl2 and 5x PTw. Kept at 4 °C overnight.

12/12/13

Ethanol washes, then first wash with glycerol and let in nutator for 3h. Exchange glycerol for glycerol+dapi and let it overnight.

13/12/13

Mounted slides of Novocrania and took pictures.