User:Karmella Haynes/Notebook/PcTF Genomics/2013/07/06: Difference between revisions
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''' | '''Luciferase activity assay - time point: 4 days'''<br> | ||
> Samples | |||
<u>Cell prep</u> | |||
* 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 | |||
<u>Assay reagents</u> | |||
* Used the Biotium Steady-Luc Firefly HTS Assay Kit (same as [http://openwetware.org/wiki/User:Karmella_Haynes/Notebook/PcTF_Genomics/2013/07/04 6/18/13]) | |||
<u>Luc assay</u> | |||
* 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 [http://openwetware.org/wiki/User:Karmella_Haynes/Notebook/PcTF_Genomics/2013/07/04 6/18/13] | |||
<u>Cell counts</u> | |||
* Wang lab's Accuri flow cytometer. | |||
* Set machine to read 20 uL of cells | |||
* Be sure to "clean" with water-run in between samples | |||
{| class="wikitable" border="0" cellspacing="3" <!-- Rxn. table --> | {| class="wikitable" border="0" cellspacing="3" <!-- Rxn. table --> | ||
|-valign="top" | |-valign="top" | ||
| <u> | | <u>Sample ID</u> || <u>Gated count/ 20 μL</u> || || <u>Cells/ 100 μL</u> | ||
|- | |- | ||
| | | sample 1 || 13,844 || x5 = || 69,220 | ||
|- | |- | ||
| | | sample 2 || 12,286 || x5 = || 61,430 | ||
|- | |- | ||
| | | sample 3 || 15,784 || x5 = || 78,920 | ||
|- | |- | ||
| | | sample 4 || 16,323 || x5 = || 81,615 | ||
|- | |- | ||
| | | sample 5 || 16,545 || x5 = || 82,725 | ||
|- | |- | ||
| | | sample 6 || 17,885 || x5 = || 89,425 | ||
|} | |} | ||
Revision as of 15:02, 6 July 2013
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07/06/13
Luciferase activity assay - time point: 4 days
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