11:00 A – 8:00P
Cell culture and cell labeling
- Start washing out direct-labeled primaries from cell-culture plates.
- Split cells, toss old plates.
- plate cells on round cover glass in 12 well pates. See if these adhere any better for FISH-on slides.
- RT incubation of tertiary antibody
- wash out tertiary
- mount embryos for confocal: no evidence of H3. H3K27ac is backgroundy. shp-dig is strong in dapi and 647, but backgroundy in cytoplasm in 647.
What keeps AbdB off (before Pc)?
Confocal: s2 cells with DFISH probes. All samples look like control, suspect cross contamination of probe between wells (whicking along lid? Maybe should do these unlidded).
- negative controls for RNA FISH: treat cells with RNase, then label
- no probe control