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 Pc-TF Genomics
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06/18/13
- Luciferase activity assay
Luciferase activity assay - time point 1
Cell prep
- Harvested cells
- Seeded new dox+ plate with 1/2 of cells
- Pelleted other half, resuspended in 2 mL FACS buffer
Assay reagents
- Used the Biotium Steady-Luc Firefly HTS Assay Kit
- Thawed reagents to room temp
- 6 experimental samples x 3 repliates = 18 assays (one time point)
- Need 100 μL complete steady-luc buffer per assay = 1800 μL complete steady-luc buffer
- Dissolved dry D-luciferin to make 50x stock by adding 80 μL dH2O to the vial
- Made 2000 μL complete steady-luc buffer: Add 40 μL 50x D-luciferin to 2 mL buffer
Luc assay
Cell count
- Wang lab's Accuri flow cytometer. Set machine to sample 2 uL of cells
| Reagent |
Volume
|
| reagent 1 |
# μL
|
| reagent 2 |
#
|
| reagent 3 |
#
|
| reagent 4 |
#
|
| dH2O |
#
|
| |
# μL
|
--> Reaction conditions
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Pc-TF Genomics
