Check PCRs (in blocks 3A (sim screening) and 3B (Espl screening)
- better way to do this: complementation test back to original sim[D] and Espl[D] lines. Just need to expand those lines back to bottles, will need ~100 vials of each
- Need to screen lines to ensure mutation is stable, not lost to double balancers. TM2/TM6 flies are ebony, others should carry candidate E/S alleles.
- DNA antibody (to think about) from Millipore. Measure DNA accessability to antibodies in heterochromatic regions?
Working on lab meeting presentation
- Figures to explain compaction assay, looking at ends
- new bottles for MTDs
- Flip fly stocks (just rack 2. Rack 1 flipped at end of Nov. Ready to flip again after group meeting)
- Discuss recombineering crosses with Sam Kunes. Recommends backcross complementation tests with original lines. Of course! This is much better than PCR mapping of breakpoints. Yay! Need to expand sim[D] and Espl[D] lines again.
- Expand Esc ? (to test after break?)
- New YW bottles (to set up cages after break)
- Move cells out of 29C incubator (Colenso has reserved for 70C)
- 750 #2 405 signal looks bad. Much stronger signal from 405-750 #1 (left most bottom). Use this one for STORM
- Both 488 750 double labeled wells no good. 488 and 405 signal very weak.
- 488 #2 no good. 488 #1 very stong. Both Cy3B wells very strong.
- switch back to 256×256 view — swap in the 50 mm focal lens
- updated thresholds for 647 and 750 STORM parameters files for Spot Counter
- TIRF angle has changed dramatically. 13.25 good TIRF.
- 647 looks very dim in STORM (more like 750), low contrast. Not sure why?
- # check 405 power
- # bead slides
coverslip Ucryo staining
- Found another coverslip that has some adhered (though only partially, no full embryo section is flat). Attempting DNA FISH + Hp1A in embryos. Prepping for DFISH.
- Make new 20% dextran-sulfate in 4x SSCT + 50% Formamide (2X SSCT final conc.)
- Sigmacote silconize slide. hybe sections on slide O/N. (Rinse off in coplin jar tomorrow).
- z-calibration files with substantial drift z-drift during calibration are impossible to use.
- Use previous passable z-fits, and run data on Cajal.
- # check with Alec about DSHB antibody order
- # abcam antibody?