User:Karmella Haynes/Notebook/PcTF Genomics/2013/07/04: Difference between revisions

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Line 45: Line 45:
| <u>Sample ID</u> || <u>Gated count/ 20 μL</u> || &nbsp; || <u>Cells/ 100 μL</u>
| <u>Sample ID</u> || <u>Gated count/ 20 μL</u> || &nbsp; || <u>Cells/ 100 μL</u>
|-
|-
| sample 1 || ### || x5 = || 54,400
| sample 1 || ### || x5 = || ###
|-
|-
| sample 2 || 10,430 || x5 = || 52,150
| sample 2 || ### || x5 = || ###
|-
|-
| sample 3 || 11,897 || x5 = || 59,485
| sample 3 || ### || x5 = || ###
|-
|-
| sample 4 || 10,316 || x5 = || 51,580
| sample 4 || ### || x5 = || ###
|-
|-
| sample 5 || 9,784 || x5 = || 48,920
| sample 5 || ### || x5 = || ###
|-
|-
| sample 6 || 11,549 || x5 = || 57,745
| sample 6 || ### || x5 = || ###
|}
|}


Revision as of 20:16, 4 July 2013

Pc-TF Genomics <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page
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07/04/13

  • Gl4-EED/luc time/dose - luc assay & cell counts
  • Happy July 4th! :)

Luciferase activity assay - time point: 4 days


Cell prep

  • Harvested cells (induced on 6/30/13)
    • Seeded new 2 new plates with non-induced cells from 6/30/13
    • 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 ### x5 = ###
sample 2 ### x5 = ###
sample 3 ### x5 = ###
sample 4 ### x5 = ###
sample 5 ### x5 = ###
sample 6 ### x5 = ###



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