Saturday 11/10/12

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)
Ordering Info
S2R+ cells
Drosophila Genomics Resource Center
$150 + 100 for shipping
M3 Insect Cell medium
For S2 and S2R+:

FBS Heat-inactivation:

  1. Prepare a water bath at 56°C.
  2. Completely thaw a bottle of FBS to room temperature.
  3. Incubate FBS in a water bath at 56°C for 30 minutes.
  4. Aliquote heat-inactivated FBS in 50 mL Falcon tubes and store at -20°C.
  5. Use one 50 mL tube of heat-inactivated FBS per 500 mL BPYE medium.


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