User:Karmella Haynes/Notebook/PcTF Genomics/2013/06/10: Difference between revisions

From OpenWetWare
Jump to navigationJump to search
Line 12: Line 12:
* Split cells: Gal4-EED/luc shipment 1 and shipment 2 T-75 flasks (1:5)
* Split cells: Gal4-EED/luc shipment 1 and shipment 2 T-75 flasks (1:5)
* Split dox induced cells
* Split dox induced cells


----
----

Revision as of 16:09, 11 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>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>


06/10/13

  • Split cells: Gal4-EED/luc shipment 1 and shipment 2 T-75 flasks (1:5)
  • Split dox induced cells


Gal4EED/luc test
> Dox dose response test


6-well plates for luc activity assays & flow cytometry, etc.

  • 6-well plate (originally plated 5/29/13), shipment 1 cells
  • Split 1:4 into 2 new 6-well plates: resuspend detached cells in a final volume of 4 mL puro+ medium, add 1 mL cells to 3mL fresh medium +proper amount of dox
Wells dox μg/mL Volume dox (1 mg/mL) in 4.0 mL medium
1 0 ---
2 0.1 0.5 μL
3 0.2 1.0 μL
4 0.5 2.0 μL
5 1.0 4.0 μL
6 2.0 8.0 μL


24-well plate for microscopy:

  • Use leftover suspension from 6-well plating
  • For each sample (1-6) add 0.5 mL suspension to 0.5 mL fresh medium +proper amount of doc in 4 wells
  • Cells will be fixed and stained directly in the plate, following David Dreher's protocol.



Retrieved from "https://openwetware.org/mediawiki/index.php?title=User:Karmella_Haynes/Notebook/PcTF_Genomics/2013/06/10&oldid=702296"