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

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(Autocreate 2013/06/10 Entry for User:Karmella_Haynes/Notebook/PcTF_Genomics)
 
(mm/dd/yy)
Line 8: Line 8:
  
  
==mm/dd/yy==
+
==06/10/13==
 
<!-- Precede finished items with a checkmark &#x2713; -->
 
<!-- Precede finished items with a checkmark &#x2713; -->
* Line item 1
+
* Split cells: Gal4-EED/luc shipment 1 and shipment 2 T-75 flasks (1:5)
 +
* Split dox induced cells
  
 
----
 
----
'''Line item 1'''<br>
+
'''Gal4EED/luc test'''<br>
> Samples
+
> Dox dose response test<br>
  
{| class="wikitable" border="0" cellspacing="3" <!-- Rxn. table -->
+
 
|-valign="top"
+
6-well plates for luc activity assays & flow cytometry, etc.
| <u>Reagent</u> || <u>Volume</u>  
+
* 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
 +
 
 +
{| class="wikitable" border="0" cellspacing="5" <!-- Cell plating table -->
 
|-
 
|-
| reagent 1 || # μL
+
| <u>Wells</u> || <u>dox μg/mL</u> || <u>Volume dox (1 mg/mL) in 4.0 mL medium</u>
 
|-
 
|-
| reagent 2 || #
+
| 1            || 0        || ---     
 
|-
 
|-
| reagent 3 || #
+
| 2            || 0.1        || 0.5 μL
 
|-
 
|-
| reagent 4 || #
+
| 3            || 0.2        || 1.0 μL
 
|-
 
|-
| dH<sub>2</sub>O || #
+
| 4            || 0.5        || 2.0 μL
 
|-
 
|-
| &nbsp; || # μL
+
| 5            || 1.0        || 4.0 μL
 +
|-
 +
| 6            || 2.0        || 8.0 μL  
 
|}
 
|}
  
--> Reaction conditions
+
 
 +
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.
  
  

Revision as of 16:09, 11 June 2013

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