9:30 A – 7:40P, 9:00P – 12:00A
Confocal dot distances
- Finish O/N confocal run of Ubx-AbdA data
Probe making
- sent order to LC Sciences for chromatin types
- On why previous probe might have failed:
In situs
- rinsing out primary antibodies
Cell culture
- Check staining. Looks promising
- blocking, will add antibodies – Dm01-405,750, H3K9me2-488
- stained
- confocal imaging — some cells have very suggestive dots, but quite few cells.
- Staining has probably failed.
Fly Work
- split MTD large cage into too (too dense, not laying sufficiently well).
- set up small cage of Su(Z)12,MTD F1s. — skipped, these embryos are starting to hatch and crawl after all. Maybe Pc1 will work better?
- collect larvae in buffer for dissection.
- # set up snail double balancing crosses generation 2
Polytene chromosome
- Dissect and mount polytene cells
- attempt mechanical chromosomal spreading. Some cells at least appear well spread, might need to repeat. Could attempt on poly-L coverglass instead of poly-L slides.
- Label with centromere stain. Left in 90C too long (30 min?). And dry.
- slide #2 labeled with DAPI, and then with H3K9me2. No staining. Dm01 works well. Try H3K27me3 tomorrow. If this fails, look up protocol?