9:30 A – 7:48 PM, 10:50 – 11:45PM
Fly work
- Flip collection plates on MTD x SCM, MTD, and sim[D], 9:30A
- Flip collection plates, sim[D] 1:15 P
- Collect virgin Act5-Gal4, MTDs, and sim/Espl. Sort sim/Espl against humoral bristles.
- New crosses (in rack at 22C)
- TRIP.Ph-p x Act5-Gal4
- TRIP.Ph-d x Act5-Gal4
- TRIP.mbt x Act5-Gal4
- TRIP.Su(z)12 x Act5-Gal4
- TRIP.Rad21 x Act5-Gal4
- TRIP.Pc1 x Act5-Gal4
- TRIP.Pc2 x Act5-Gal4
- TRIP.Pc3 x Act5-Gal4 (- cntrl, viable in MTD xs)
- TRIP.Scm x Act5-Gal4 (+ cntrl, lethal in MTD xs)
- Moved all 12 Espl/sim x TM2/TM6 crosses to 22C to grow.
- Freeze MTD x PSC and MTD x PC3
- Fix embryos (3PM)
- Fix embryos (7PM) — [skipped]
- PM virgin collection: Act5-G4 and MTDs.
Western digital MyBook studio 2 refuses to format for windows.
CD software freezes at ‘Preparing’ when attempting to install on Tuck or Monet.
Copying data to GRAID
- Meeting with Eric Wang,
- discuss SR imaging, recommend HCBI ELYRA to try out stuff and start contact in official channels to see what options are there.
- discuss RCA on ligated tethered antibodies for colocalization.
Cell culture DNA-FISH test
- toll, y, antp, ubx, y-2, abdA-2, antp2, cent.
- Added 50 uL + of DFISH hybe soln to compensate for evaporation…
Embryo DNA-FISH: day 2 antibody labeling. (4C O/N).
Probe making
- Test get of DFISH probes from BAC5 and 1 (Purple labels), BAC 5 -red label at 4:1 mix:polI.
- Probes look okay, large smear from 100bp-5kb. Ask Fred about getting this narrower? Use more DNAse?
- concentration (with dye in buffer, relative to ddH2O blank) is 220 ng/mL = pretty good? (despite rather faint on typhoon? can see well on gel box. Should dilute ladder another 10x at least).
Sectioning: cut 4 coverslips of sample 1- inv,tou-dig-Cy3B, Epc-514. see how it goes. Except need to fix steve for different camera position.
Francis lab group meeting — discuss snoRNA paper.