User:Karmella Haynes/Notebook/Polycomb project/2010/08/02

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  • ✓ H3me Reporter clone screening: cells are too crowded for transfection, split 1:4 for transfection tomorrow (12-well plates, ab-free, 1 mL per well); make back-ups in 6-well plates (FTRx-hygro maintenance medium)
  • ✓ KAH131 screening: check RFP expression, expand selected clones
  • ✓ Thaw: KAH126-3 for repeat of senescence expt.
  • ✓ Microscopy: finish Pc-ATF images if DAPI and RFP signal still good

KAH131 screening
> RFP expression -dox/ vs. +dox

  1. KAH131-4 (+/++)
  2. KAH131-8 (+/++)

--> Note: These two and four other RFP+ clones showed lots of cell death after dox induction (~4 days); FlyPc-RFP-VP64 appears to be highly toxic
--> Expand two lines in 6-well plates (FTRx+puro maintenance medium)
--> Might omit this line from further studies if difficult to grow