Beginning a double in situ with en dnp…


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Beginning a double in situ with:

en (dnp) + gli (dig)
en (dnp) + fgf8/17/18 (dig)

02/06/13

Added probes.

04/06/13

First washing day normal. Washes followed at times, but second 0.2x SSC took 40 instead of 20 min. Added Anti-Dig-POD at 1:250 concentration.

05/06/13

Second washing day normal.

06/06/13

Developed first probe. CAOS after detergent wash, during subsequent PTw washes the tape glued to the cover and FLIPPED the 4-well dish… I recovered most embryos and many remained in their wells, but for sure there has been some mixing. I added the recovered embryos to a third well.

Apart from that the first probe works. And incubated with Anti-DNP at 1:100.

07/06/13

Regular washes.

08/06/13

Developed second probe and let it in PTw in the cold room until 12/06/13.

12/06/13

DAPI staining for 20 min, one wash of PTw and glycerol overnight.