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.