User:Karmella Haynes/Notebook/miR Trigger/2009/12/06

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12/06/09

 * &#x2713; Transfection: KAH93, 94, 95/MV2 (RFP); KAH99, 100, 101/MV2 (CFP)

Transfections > U2OS cells + Gal4DB-PEST-RFP constructs > 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. Cells in 6-well plates too crowded, discard; do same-day plating/ transfection from back-up stocks > Harvest cells, plate 5x105 cells/3.5 ml ab-free medium in 6-well plates > Add 500 μL complexes to each well (4 ml med. each); Grow cells at 37&deg;C > Check RFP & CFP expression (microscopy) after 18-24 hours


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