9:45 A – 7:30P, 8:45P-12:30A
Embryo collection
- Flip ESC embryos
- Flip all other embryos (w/o yeast, just as a holding pattern).
- check vitality — Psc1 has a 5 of 100s hatched, others have 0 hatched yet. Will give them another 24 hrs.
- Fix Esc embryos 1:45P. (8:00P-9:30A collection). 5-18 hr collection
- All cages moved to RT (incubator set to 42C for O/N hybe)
Group meeting
Confocal
- Esc Ubx + AbdB staining: looks excellent, should definitely screen other lines
Embryo Staining
- Screening mutants for Ubx and/or Abd-B mis-expression
- scheduled Confocal time tonight to image
- Mat alpha tubs mostly infertile? (no DAPI?!) refix tomorrow and recheck.
- strong ubiquitous AbdB expression + morphological changes in all zygotic mutants checked.
- Ubx stain not very strong
Embryo U-cryo-section Staining
- labeling tissue fragments with Ubx or Abd-B and post-fix prior to BXC labeling
- Ubx coverslip on good embryo sections
- AbdB coverslip one solid patch of lots of embryos, lots of ages.
- O/N incubation in BX-c probe (1 uL primary 1 uL secondary in 50 uL hybe)
Other
- chat with Jeff about oligoPAINTs. Plan to do emPCR together on Thursday
- confirm oligoPAINT primers ready to go for emPCR (in DFISH box 2)
- # discussion with new G0 student
- Order new formaldyhde from Polysciences using PO. Price went up $6.00, hopefully the PO still goes through?
- Attempted to order new heptane from Fisher with PO — item prohibited (see email to Alec).