Difference between revisions of "User:Karmella Haynes/Notebook/PcTF Genomics/2013/06/13"

From OpenWetWare
Jump to: navigation, search
(Autocreate 2013/06/13 Entry for User:Karmella_Haynes/Notebook/PcTF_Genomics)
 
(mm/dd/yy)
Line 8: Line 8:
  
  
==mm/dd/yy==
+
==06/13/13==
 
<!-- Precede finished items with a checkmark &#x2713; -->
 
<!-- Precede finished items with a checkmark &#x2713; -->
* Line item 1
+
* Gal4-EED/luc cells IFC - secondary stain & imaging
 +
 
  
 
----
 
----
'''Line item 1'''<br>
+
'''Gal4-EED/luc cells IFC'''<br>
> Samples
+
* Samples: first row of wells in the 24-well plate.
 +
* Keep all others under PBS
 +
 
 +
# 0 μg/mL dox
 +
# 0.1
 +
# 0.2
 +
# 0.5
 +
# 1.0
 +
# 2.0
 +
 
 +
* Follow standard protocol; wash 3x with 1xPBS
 +
* Use 1:1000 dilution of anti-rabbit Alexa488 and 1:1000 Hoescht (in 15 μL Gentex horse serum blocking solution per well)
 +
* Incubate under parafilm "covers" for one hour at room temperature in light-tight box with wet paper towels
 +
* Use GFP setting on Nikon scope to record images
  
{| class="wikitable" border="0" cellspacing="3" <!-- Rxn. table -->
+
Results:
|-valign="top"
+
* Saw some cells with very bright signal, some dim in 0 μg/mL dox well
| <u>Reagent</u> || <u>Volume</u>
+
* Signal appeared to be dimmer overall for all others.
|-
 
| reagent 1 || # μL
 
|-
 
| reagent 2 || #
 
|-
 
| reagent 3 || #
 
|-
 
| reagent 4 || #
 
|-
 
| dH<sub>2</sub>O || #
 
|-
 
| &nbsp; || # μL
 
|}
 
  
--> Reaction conditions
 
  
  

Revision as of 17:46, 17 June 2013

Owwnotebook icon.pngPc-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>


06/13/13

  • Gal4-EED/luc cells IFC - secondary stain & imaging



Gal4-EED/luc cells IFC

  • Samples: first row of wells in the 24-well plate.
  • Keep all others under PBS
  1. 0 μg/mL dox
  2. 0.1
  3. 0.2
  4. 0.5
  5. 1.0
  6. 2.0
  • Follow standard protocol; wash 3x with 1xPBS
  • Use 1:1000 dilution of anti-rabbit Alexa488 and 1:1000 Hoescht (in 15 μL Gentex horse serum blocking solution per well)
  • Incubate under parafilm "covers" for one hour at room temperature in light-tight box with wet paper towels
  • Use GFP setting on Nikon scope to record images

Results:

  • Saw some cells with very bright signal, some dim in 0 μg/mL dox well
  • Signal appeared to be dimmer overall for all others.