User:Karmella Haynes/Notebook/miR Trigger/2009/12/04

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12/04/09

  • Transfection: KAH93, 94, 95/MV2 (Plasmids not yet confirmed, postpone)

Transfections
> U2OS cells + Gal4DB-PEST-RFP constructs
> Use Lipofectamine, 6-well format

Wells Plasmid DNA Volume Fugene Opti-MEM (total)
1 KAH93/MV2 2 μg ## μL 4 μL 500 μL
2 KAH94/MV2 " ## μL " "
3 KAH95/MV2 " ## μL " "
4 KAH36(control) " ## μL " "
5 mock --- --- " "
6 untreated --- --- --- "

> 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.
> Add 500 μL complexes to each well (4 ml med. each); Grow cells at 37°C
> Assay RFP after 18-24 hours



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