9:40 A – 7:45P, 9:00P – 1:00A
Dfd/tub analysis
- Break down O/N confocal run
- 3 channels, speed 6, 22 positions, 17.5 finished in 11 hour run. contact zeiss rep Casey about changing channels between z-stacks rather than every section, should be much faster.
- annotating ages from overview images of last nights’ Dfd tub data
- Analyzing data (see post).
Probe making,
- spin down dissolving DNA from pulverized gels, remove supernatant for precipitation, return remaining gel to 55C shaker with fresh ddH2O to elute remaining probe.
- precipitate eluted probe
In situs Day 2
- hot washes
- block
- primary antibodies O/N
Ucryo sections on coverglass
- dk a-rb-488 doesn’t label either, so hopefully not a problem with the abcam dk a-rab 750 I made earlier.
- reblocked and staining with H3K9me2 again (5:300 of the rediluted in glycerol stock). It would be really nice if this would work.
Cell culture test 50kb probe
- Plate new cells from overgrown 11/20 culture flask
- Odd colored precpitate observed in mixing flask. Cells tossed.
- Contact Nicole about starting a new culture going from frozen stock. Current line has been in continuous culture since Aug.
- Fix new plated cells. (Attachment was poor / or even new small squeeze bottle has too much force, most cells detached).
- Prehybe 60C, denature probe, denature cells + probe at 92, incubate 37C O/N