User:Karmella Haynes/Notebook/PcTF Genomics/2013/06/15

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(Autocreate 2013/06/15 Entry for User:Karmella_Haynes/Notebook/PcTF_Genomics)
Current revision (23:11, 26 June 2013) (view source)
(06/15/13)
 
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==mm/dd/yy==
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==06/15/13==
<!-- Precede finished items with a checkmark &#x2713; -->
<!-- Precede finished items with a checkmark &#x2713; -->
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* Line item 1
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* Gal4-EED/luc Luciferase ICC - fixation, permeabilization, primary ab staining
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* Gal4-EED/ luc dox induction - shipment 2 cells
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* PcTF transfection for U2OS, outgrowth #1
 +
 
----
----
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'''Line item 1'''<br>
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'''Gal4-EED/luc Luciferase ICC'''<br>
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> Samples
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{| class="wikitable" border="0" cellspacing="3" <!-- Rxn. table -->
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Samples (Shipment 1 cells)
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|-valign="top"
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# 0 μg/mL dox
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| <u>Reagent</u> || <u>Volume</u>
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# 0.1
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|-
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# 0.2
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| reagent 1 || # μL
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# 0.5
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|-
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# 1.0
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| reagent 2 || #
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# 2.0 (induced on 6/02/13, 13 days)
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|-
+
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| reagent 3 || #
+
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|-
+
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| reagent 4 || #
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|-
+
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| dH<sub>2</sub>O || #
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|-
+
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| &nbsp; || # μL
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|}
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--> Reaction conditions
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* Followed standard protocol
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* Harvested cells from one 6-well plate (split plate #2 for further analyses)
 +
* At the primary antibody staining step, keep samples and reagents on ice overnight in the cold room
 +
 
 +
 
 +
----
 +
'''Gal4-EED/luc dox induction'''<br>
 +
 
 +
Samples (Shipment 2 cells); new 6-well plate...
 +
# 0 μg/mL dox
 +
# 0.1
 +
# 0.2
 +
# 0.5
 +
# 1.0
 +
# 2.0 (induced on 6/02/13, 13 days)
 +
 
 +
* For this set, test earlier time points and try luc activity assay
 +
 
 +
 
 +
 
 +
----
 +
'''PcTF transfection for U2OS, outgrowth #1'''
 +
* 6-well plates: 1-3 = +PcTF, 4-6 = mock (lipo only)
 +
* Expand cells from 6-well plates in individual 10 cm plates (10 mL medium). 6 10 cm plates total.
 +
* Use standard growth medium.
 +
* Use all cells. Do not dilute.

Current revision

Pc-TF Genomics Main project page
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06/15/13

  • Gal4-EED/luc Luciferase ICC - fixation, permeabilization, primary ab staining
  • Gal4-EED/ luc dox induction - shipment 2 cells
  • PcTF transfection for U2OS, outgrowth #1



Gal4-EED/luc Luciferase ICC

Samples (Shipment 1 cells)

  1. 0 μg/mL dox
  2. 0.1
  3. 0.2
  4. 0.5
  5. 1.0
  6. 2.0 (induced on 6/02/13, 13 days)


  • Followed standard protocol
  • Harvested cells from one 6-well plate (split plate #2 for further analyses)
  • At the primary antibody staining step, keep samples and reagents on ice overnight in the cold room



Gal4-EED/luc dox induction

Samples (Shipment 2 cells); new 6-well plate...

  1. 0 μg/mL dox
  2. 0.1
  3. 0.2
  4. 0.5
  5. 1.0
  6. 2.0 (induced on 6/02/13, 13 days)
  • For this set, test earlier time points and try luc activity assay



PcTF transfection for U2OS, outgrowth #1

  • 6-well plates: 1-3 = +PcTF, 4-6 = mock (lipo only)
  • Expand cells from 6-well plates in individual 10 cm plates (10 mL medium). 6 10 cm plates total.
  • Use standard growth medium.
  • Use all cells. Do not dilute.



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