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

From OpenWetWare

< User:Karmella Haynes | Notebook | PcTF Genomics | 2013 | 06(Difference between revisions)
Jump to: navigation, search
(06/15/13)
Current revision (23:11, 26 June 2013) (view source)
(06/15/13)
 
(6 intermediate revisions not shown.)
Line 11: Line 11:
<!-- Precede finished items with a checkmark &#x2713; -->
<!-- Precede finished items with a checkmark &#x2713; -->
* Gal4-EED/luc Luciferase ICC - fixation, permeabilization, primary ab staining
* Gal4-EED/luc Luciferase ICC - fixation, permeabilization, primary ab staining
 +
* Gal4-EED/ luc dox induction - shipment 2 cells
 +
* PcTF transfection for U2OS, outgrowth #1
Line 16: Line 18:
'''Gal4-EED/luc Luciferase ICC'''<br>
'''Gal4-EED/luc Luciferase ICC'''<br>
-
Samples
+
Samples (Shipment 1 cells)
# 0 μg/mL dox
# 0 μg/mL dox
# 0.1  
# 0.1  
Line 22: Line 24:
# 0.5
# 0.5
# 1.0
# 1.0
-
# 2.0
+
# 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)
* 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
* 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
Previous entry      Next entry


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.



Personal tools