User:Karmella Haynes/Notebook/Polycomb project/2009/12/21

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12/21/09

 * &#x2713; Transfections: Polycomb ATF's for stable lines

Transfections > U2OS Flp-in T-REx cells + Polycomb-ATF plasmids (puro) > Use Lipofectamine, 6-well format

> Add Lipo to 250 μL Opti-MEM --> R.T/ 5 min. > Add DNA to 250 μL Opti-MEM > Add DNA mix to Lipo mix --> R.T./ 20 min. > Remove 500 μL medium from each well; Add 500 μL complexes to each well (4 ml med. each); Grow cells at 37&deg;C > Refresh medium after 5 hours (ab-free) > Grow for two days, then plate dilution cultures in selection medium (puro)


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