User:Karmella Haynes/Notebook/PcTF Genomics/2013/06/19: Difference between revisions

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(Autocreate 2013/06/19 Entry for User:Karmella_Haynes/Notebook/PcTF_Genomics)
 
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----
'''Line item 1'''<br>
'''Luciferase activity assay - time point 2'''<br>
> Samples
 
 
<u>Cell prep</u>
* Harvested cells
** Seeded new dox+ plate with 1/2 of cells
** Pelleted other half, resuspended in 2 mL FACS buffer
 
 
<u>Assay reagents</u>
* Used the Biotium Steady-Luc Firefly HTS Assay Kit (same as 6/18/13)
 
 
<u>Luc assay</u>
* Filtered '''700 μL cells''' through strainer caps
* Other steps same as 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>Reagent</u> || <u>Volume</u>  
| <u>Sample ID</u> || <u>Gated count/ 20 μL</u> || &nbsp; || <u>Cells/ 100 μL</u>
|-
|-
| reagent 1 || # μL
| sample 1 || || x5 = ||
|-
|-
| reagent 2 || #
| sample 2 || || x5 = ||
|-
|-
| reagent 3 || #
| sample 3 || || x5 = ||
|-
|-
| reagent 4 || #
| sample 4 || || x5 = ||
|-
|-
| dH<sub>2</sub>O || #
| sample 5 || || x5 = ||  
|-
|-
| &nbsp; || # μL
| sample 6 || || x5 = ||  
|}
|}
--> Reaction conditions




Revision as of 21:37, 19 June 2013

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mm/dd/yy

  • Line item 1

Luciferase activity assay - time point 2


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 (same as 6/18/13)


Luc assay

  • Filtered 700 μL cells through strainer caps
  • 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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