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 Pc-TF Genomics
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07/06/13
- Gal4-EED/luc dox time point (48 hours)
- Gal4-EED/luc PcTF transfection - microscopy & flow cytometry
Luciferase activity assay - time point: 2 days
Cell prep
- Harvested cells (induced on 7/04/13)
- Seeded new 2 new plates with non-induced cells from 7/04/13
- Pelleted all of the dox+ cells, resuspended in 2 mL FACS buffer
Assay reagents
- Used the Biotium Steady-Luc Firefly HTS Assay Kit (same as 6/18/13)
Luc assay
- Filtered 700 μL cells through strainer caps
- Used opaque white Costar plate (from Rege lab)
- Samples loaded in triplicate (by columns)
- Included luc buffer + FACS-buffer "blank" sample (well D1)
- Other steps same as 6/18/13
Cell counts
- Wang lab's Accuri flow cytometer.
- Set machine to read 20 uL of cells
- Be sure to "clean" with water-run in between samples
| Sample ID |
Gated count/ 20 μL |
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Cells/ 100 μL
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| sample 1 |
### |
x5 = |
###
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| sample 2 |
### |
x5 = |
###
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| sample 3 |
15,784 |
x5 = |
78,920
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| sample 4 |
16,323 |
x5 = |
81,615
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| sample 5 |
16,545 |
x5 = |
82,725
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| sample 6 |
17,885 |
x5 = |
89,425
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Pc-TF Genomics
