User:Karmella Haynes/Notebook/Polycomb project/2010/10/27

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10/27/10

 * &#x2713; Transfections: HEK293 Gal4EED + Pc-ATF's

Transfections > Try transfecting with Pc-ATF first, then inducing H3K27me3 > HEK293 Gal4-EED 23;4;9 cells, no dox induction > Use Fugene, 12-well format, 1.5 mL medium per well > 2 plates total (1 control, 1 for dox induction time course)

> Master mix (2x, 1 per plate): 3 μL Fugene + 97 μL Opti-MEM --> R.T./ 5 min. > Add 2x vol. DNA --> R.T./ 20 min. > Remove 0.5 mL medium from each well (decrease volume to improve transfection efficiency) > Add 50 μL DNA/Fugene mix to each well (1.5 ml medium/ well); Grow cells at 37&deg;C > Check RFP after 18-24 hours


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