User:Karmella Haynes/Notebook/PcTF Genomics/2013/06/18: Difference between revisions
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* Need 100 μL complete steady-luc buffer per assay = '''1800 μL complete steady-luc buffer''' | * Need 100 μL complete steady-luc buffer per assay = '''1800 μL complete steady-luc buffer''' | ||
* Dissolved dry D-luciferin to make 50x stock by adding 80 μL dH<sub>2</sub>O to the vial | * Dissolved dry D-luciferin to make 50x stock by adding 80 μL dH<sub>2</sub>O to the vial | ||
<u>Luc assay</u> | <u>Luc assay</u> | ||
* Filtered 500 μL cells through strainer caps (<font color="red">should use 700 μL next time</font>) | |||
* Transferred 100 μL strained cells to 3 wells in 96-well plate (black walls; clear, flat bottom) | |||
** Saved remainder of strained cells for flow cytometry/ counting | |||
* Made 2000 μL complete steady-luc buffer: Added 40 μL 50x D-luciferin to 2 mL buffer | |||
* QUICKLY Added 100 μL of complete steady-luc buffer to each sample (pipetted up/down once to mix) | |||
* Incubated at room temp/ 10 min. | |||
<u>Cell count</u> | <u>Cell count</u> |
Revision as of 18:35, 18 June 2013
Pc-TF Genomics | <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page <html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>Previous entry<html> </html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html> | ||||||||||||||
06/18/13
Luciferase activity assay - time point 1
--> Reaction conditions
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