10:45 A – 8:00P
Snail project stuff
- Collect virgin Pr/Tm3Z, Sp/CyOZ, and TM2/TM6 (males + virgins)
- Order primers for screening E(spl) and sim[D]. Also for en and inv introns
DNA FISH probe design
- Wrote new matlab script for concatinating text files (e.g. .wig files), for easier, faster import into UCSC genome browser
- Designing tiling sets of primers for tou gene to try Fred’s method of template making for nicked translation. (~40 min for 10 sets, should write a short program to do this if we intend to do much more this way).
- Installing python scripts from OligoPaints project
- Step 1: make BED files for each region of interest. (actually just need numbers)
- GREEN region 1073 probes 9.6 probes/kb (not too dense — most green regions on 2R 6-9 probes/kb).
- YELLOW region 1725 probes 17 probes/kb
- BLUE region 1815 probes, 15/kb
- BLACK 1625 14.5 probes/kb
- Mapped region
Order H3K9me2. Millipore image looks the most punctate. (Abcam and ActiveMotif look okay as well).
DNA FISH S2 cells on coverglass — confocal report
- weak background staining particular to DAPI depleted nuclear regions with Toll2 probe.
- no staining at all with sxl2 probe
Cell Culture Planning
Switch to S2R+ Cell culture (adherent S2 line) ?
Medium = M3+BPYE + 10% FCS (from DGRC)
Drosophila Genomics Resource Center
$150 + 100 for shipping
M3 Insect Cell medium
For S2 and S2R+:
- Prepare a water bath at 56°C.
- Completely thaw a bottle of FBS to room temperature.
- Incubate FBS in a water bath at 56°C for 30 minutes.
- Aliquote heat-inactivated FBS in 50 mL Falcon tubes and store at -20°C.
- Use one 50 mL tube of heat-inactivated FBS per 500 mL BPYE medium.