User:Karmella Haynes/Notebook/PcTF Genomics/2014/11/15

Pc-TF Genomics

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11/15/14 Transfection: KAH126/MV2 into U2OS Transfection: KAH126/MV2 into U2OS Strategy: instead of trying to increase transfection efficiency, do a "quasi-subcloning" where transfected cells are separated into smaller wells, and wells showing highest RFP will be selected, pooled, and re-plated. 6-well plate seeded on 11/14/14 (~2.5E5 cells/well) Change medium to p/s-free before transfection (4 mL per well) Use standard Lipofectamine LTX procedure w/ 2000 ng plasmid KAH126/MV2 mock no transfection Grow overnight, look for RFP next day Next day: harvest transfected cells and use 1/2 (5.0E5) for 2 different plates 24-well plate - ~2.1E4 cells/well 96-well plate - ~5.2E4 cells/well