User:Karmella Haynes/Notebook/Polycomb project/2010/02/11

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02/11/10

  • ✓ HEK293 #4 Luc transfection: luc assay
  • RNA preps: U2OS Flp-in T-REx (+/- dox, 2 days) Instead, include as neg. ctrl. for long-term dox induction
  • ✓ Polycomb-ATF lines: make frozen stocks from confluent plates



Luc assay
--> Promega Bright-glo kit
> Resuspend cells in 400 μL 1xPBS (cells detach easily w/o trypsin)
> Transfer 3x 100 μL to 3 wells in an opaque white 96-well plate for luc reading
> Add 10 μL to 10 μL trypan blue for cell counting (luc normalization)


> Cell count x 2 x 10,000

  1. 23 = 4.6 x 105
  2. 37 = 7.4 x 105
  3. 38 = 7.4 x 105
  4. 26 = 5.2 x 105
  5. 40 = 8.0 x 105
  6. 26 = 5.2 x 105
  7. 34 = 6.8 x 105
  8. 36 = 7.2 x 105



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