9:20 A – 6:30P, 8:00P-11:00P (remote)
S2 cell DNA-FISH labeling
- Fixing S2 cells for DFISH (1/2x PBT 5% formaldehyde)
- cells still attached, looks promising.
- cells preincubated at 92C for 3min, 60C for 20min, moved to RT
- probes denatured at 92C for <1 min (should be single stranded centromere probes denature not as necessary except for some secondary structure).
- probes added to samples at RT, samples in humidity box in 37C room.
S2 cell RNA-FISH labeling
- Fixing S2 cells for RFISH (1/2x PBT 5% formaldehyde)
- cells in prehybe at 55C downstairs (in humidity box).
- added Epc-dig, tou-bio, top row 150uL each
- added Epc-dig, en-bio, bottom row 150 uL each
- forgot to heat denature probe secondary structure. we’ll see how much this matters.
Embryo prep for STORM continued
- embryo labeling
- washing out secondary antibodies from O/N embryo staining.
- Pc embryos on final rinses
- embryos in tertiary antibodies at RT, 12:00P.
- mount Pc embryos for confocal (1:3oPM)
- post fix Pc embryos
Fly Work
- Espl x sim[D] pupa just beginning to darken. (start collecting virgins in ~2 days?)
- TRiP knockdowns just beginning to emerge. (Maybe ready for cages by Friday).
Other prep
- flipped fly cage
- made new DFISH fix mix. Aliquot and Freeze.
- Microscopy seminar: confocal
- Write to Ting about S2 cell DFISH
- Working on BJ review
- Write to Mike about kinetics paper.
- Write to Jacques, draft response to Mike.
- split S2 cells
- Book 780 ‘scope time: Thur night, Fri night, Sat night and Mon night.