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

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02/11/10

 * &#x2713; 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
 * &#x2713; 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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