Membranipora in situ and Novocrania wnt1

Starting in situ in Membranipora with dorsoventral genes and additional genes that we backgroundish.

wnt1 dlx bmp2/4 chordin fgf
nanos vasa piwi1 piwi2 pl10
Na wnt1

Standard in situ protocol with 10 min protk. Washes after glycine were slightly shorter and fixing went for 1:30h. Used 83°C for fosfatase step and prehybe at 67°C.

09/05/15

Added probes just after synthesis.

11/05/15

Novocrania were dissolved. The rest were fine. Continued with hybe washes and added anti-dig-ap.

12/05/15

Antibody washes. Plus developing. dlx and piwi1 were stopped after a couple of hours. The remaining were developed overnight at 4 °C.

13/05/15

Exchanged the AP and stopped the reaction around 1200. Did ethanol washes and left in 70% glycerol + dapi overnight at room temp.