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

From OpenWetWare
Jump to: navigation, search
(mm/dd/yy)
(fix raw html notebook nav)
 
(2 intermediate revisions by one other user not shown)
Line 2: Line 2:
 
|-
 
|-
 
|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;">Pc-TF Genomics</span>
 
|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;">Pc-TF Genomics</span>
|style="background-color: #F2F2F2" align="center"|<html><img src="/images/9/94/Report.png" border="0" /></html> [[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>[[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]]<html>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>}}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]]<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>}}
+
|style="background-color: #F2F2F2" align="center"|[[File:Report.png|frameless|link={{#sub:{{FULLPAGENAME}}|0|-11}}]][[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|[[File:Resultset_previous.png|frameless|link={{#lnpreventry:{{FULLPAGENAME}}}}]][[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]]&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;}}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]][[File:Resultset_next.png|frameless|link={{#lnnextentry:{{FULLPAGENAME}}}}]]}}
 
|-
 
|-
 
| colspan="2"|
 
| colspan="2"|
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
 +
  
 
----
 
----
Line 42: Line 43:
 
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.
  

Latest revision as of 22:45, 26 September 2017

Owwnotebook icon.pngPc-TF Genomics Report.pngMain project page
Resultset previous.pngPrevious entry      Next entryResultset next.png


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.