User:Karmella Haynes/Notebook/PcTF Genomics/2013/06/10

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(06/10/13)
Current revision (23:19, 12 June 2013) (view source)
(06/10/13)
 
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24-well plate for microscopy:  
24-well plate for microscopy:  
* Use leftover suspension from 6-well plating
* 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
+
* For each sample (1-6) add 0.5 mL suspension to 0.5 mL fresh medium +proper amount of dox in 4 wells
* Cells will be fixed and stained directly in the plate, following David Dreher's protocol.
* Cells will be fixed and stained directly in the plate, following David Dreher's protocol.

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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 dox in 4 wells
  • Cells will be fixed and stained directly in the plate, following David Dreher's protocol.



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