User:Karmella Haynes/Notebook/Polycomb project/2011/03/02

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03/02/11

  • ✓ HEK luc-repression/ Pc-TF transfection: luc activity assay


Luc activity assay
> Cells: dox-induced Gal4-EED expression/ 2 days, followed by transfection for 2 days
1-6. KAH160/MV1 (human Pc-TF), 0, 50, 100, 200, 400, 800 ng plasmid
7-12. KAH160/MV1 (human Pc-TF), 0, 50, 100, 200, 400, 800 ng plasmid

> Sample processing:

  • Discard 400 μL of the growth medium, resuspend cells in remaining ~600
  • Aliquot 3x 100 μL samples into 96-well plate
  • Save the rest for cell count (Millipore Scepter)
  • Normalize each value to cell count



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