9:35 A – 7:30, 9:30 – 10:50 P
- wash out O/N antibodies
- Flip fly collection plates 10:10A
- Request more time to write review
- Flip expanding Gal4-Bcd3’UTR lines: growing slowly only 2 bottles worth
- O/N collection plates of en-GFP, Bcd-GFP, Pc-GFP and eve-GFP into new bottles. Bcd not laying well.
- Did not heat embryo to 92C, just probe. Probably needed to heat embryos to denature DNA for hybe.
- en-GFP no obvious staining. embryos are hets, need to check more. Strong 488 background from gut
- Pc-GFP embryos look weird mostly older, not all devitelinized. Possibly a fixation issue. Try again take more care with fix and stain with anti-GFP.
- Hp1a looks very good, H3 somewhat weak, not sure why–maybe antibody washes out during multi-day treatement? Try post-fixing?
- No staining with oligo-probes. Try heat-denaturing embryos (started 5P)
- Fix Pc-GFP, eve-GFP and en-GFP embryos 1.5 – 7 hr collection
- stain these GFP lines with endogenous antibody at 4C O/N
- mixed up K27 647 and 750 antibodies upon removal, no good for reuse. Tossed. Need to make new labeled antibodies.
- K27 PcG embryos all in 100% EtOH O/N