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

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(Autocreate 2013/06/13 Entry for User:Karmella_Haynes/Notebook/PcTF_Genomics)
 
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==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

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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.




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