|
07/06/13
- Gal4-EED/luc dox time point (48 hours)
- Gal4-EED/luc PcTF transfection - microscopy & flow cytometry
- SK-N-SH +PcTF - TRIzol prep #1
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 |
|
Cells/ 100 μL
|
| sample 1 |
37,629 |
x5 = |
|
| sample 2 |
34,580 |
x5 = |
###
|
| sample 3 |
40,096 |
x5 = |
|
| sample 4 |
46,599 |
x5 = |
|
| sample 5 |
46,491 |
x5 = |
|
| sample 6 |
46,254 |
x5 = |
|
|
Pc-TF Genomics
