Sunday 11/04/12

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).

To try:

  • negative controls for RNA FISH: treat cells with RNase, then label
  • no probe control
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