8:30 A – 9:50 P
Fly work
- Flip collection plate 8:30A
- Fixing embryos 11:30A. (20 min 8% FA reaction)
- Flip all polycomb mutants into bottles
STORM
- made new bead slide, 1:25K
- imaging 2 rounds of beads for 3D fitting
- K9me2 750 in Immuno first vs. FISH first
Probe making
- Run denaturing gel to extract 500 kb probe.
- gel runs slightly crooked. cut out lane, post-stained for 10 min with 15 uL GelGREEN in a pippett tip box lid with gel running buffer. Imaged on Typhoon. destained 10 min in ddH2O, extracted using Brian’s method of O/N 55C shaker in 600 uL of .4 M ammonium acetate. Why has the ammonium acetate conc been dropped? Less volume of ddH2O in added solution?
Confocal
- Imaging whole embryo MP08 (snail minus distal) and MP12 (snail BAC controls).
- For figures for bistability