User:Karmella Haynes/Notebook/Polycomb project/2011/03/05

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03/05/11

  • ✓ Luc assay: 2-day luc-silenced cells, 4 days Pc-TF expression



Luc Activity Assay

Luc activity assay > Cells: dox-induced Gal4-EED expression/ 2 days, followed by transfection for 4 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:

  • Resuspend cells in total 1 mL medium
  • Aliquot 3x 100 μL samples into 96-well plate
  • Save the rest for cell count (Millipore Scepter)
  • Normalize each value to cell count

> Note: May have used too little Pc-TF DNA before. I think I added only 2x DNA to the 4x volume before doing serial dilutions. Should repeat this experiment again later if 800 ng result is needed.