Novocrania in situ wnts


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Another in situ now using the newly synthesized probes.

06/11/13

Added Probes.

08/11/13

First washing day. Diluted 0.2 µL of Anti-DIG-AP in 1x blocking buffer and let overnight in cold room.

09/11/13

Washing off antibody. And started developing. Exchanged the substrate 4 times and no signal. Let overnight in cold room.

10/11/13

Wnt1 signal is visible! Ring around blastopore in gastrula and posterior ventral in later stages. No bands! Continued developing until 14h, then 4°C until 18h when I exchanged the substrate and let overnight again. Wnt5 had signal in the chaetae sacs.

11/11/13

Checked and signal looks a bit stronger so I developed 40 min more and stopped the reaction. Crystals were forming in the wnt5 well. Ethanol washes and will put them in glycerol with DAPI.

12/11/13

Mount and take pictures.

Wnt1

Wnt5