10:00A – 10:30P
(8:30-10A lab exercise/training)
Bistability
- First draft of deterministic model figure and corresponding text
- Trying to figure out correct integral to compute an energy surface from the vector field. Not working so far.
Fly work
- collect virgins:
- (now have 4 vials of sim[D] and Espl[D], should be enough for 2x females a piece to cross for complimentation testing).
- 1 new cross of DB hb-LacZ
- 3 new crosses of Esc[6]/CyO x Sp/CyO
- new YW collection cage, low number of flies, hopefully works out alright.
Communication
- Contact Ajaz about concavlin A protocol
- should hear from Stas on Monday about review
- Contact Carl about issues with divergence for vector field of bistable system
- Need to integrate vector field
PCR of 500 kb probe from 3R. Freezing O/N to precipitate.
Cell culture protein DNA multiplex:
- Goals
- HP1 centromere multiplex
- evaluate 750 stain and test new 750 approach
- compare protein first + post-fix (short and longer PFA/GA treatment) to
DFISH first then protein, no post-fix (less background?)
- Coating coverslips with polyL-lysine from Eileen (30 min, under UV light). Warming up culture media.
- Fix (15 min 5% PFA), 2x PBT rinse, 15 min NaBoH (in ddH2O) treat. Block 45 min
- left half of wells abcam m-H3K9me2, right half in m-HP1 (new aliquot) at 1:200 dilution of 4C stock
- O/N incubation in PBT wash
- O/N incubation of chamber 2 in FISH probes:
- Cy5-AACAC
- Cy5-AATAT