User:Karmella Haynes/Notebook/miR Trigger/2011/02/06

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01/06/11

  • ✓ Transfections: miR sensors into U2OS plain and JDS33-47



Lipo transfection
> 2 Plates

  1. U2OS plain
  2. JDS33-47
Wells Plasmid DNA Volume Lipo Opti-MEM (total)
1 KAH179/176/MV8 2 μg 7.3 μL 4 μL 500 μL
2 KAH180/176/MV8 " 9.4 μL " "
3 KAH179/177/MV8 " 4.8 μL " "
4 KAH180/177/MV8 " 6.4 μL " "
5 KAH179/178/MV8 " 5.1 μL " "
6 KAH180/178/MV8 " 4.8 μL " "

> 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.
> Plate ~2.5 x 105 cells in each well; use 3.5 mL medium
> Add 500 μL complexes to each well (4 ml med. each); Grow cells at 37°C
> Change to plain medium and grow for 3 days



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