Difference between revisions of "User:Karmella Haynes/Notebook/PcTF Genomics/2013/06/15"

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|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>}}
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|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}}}}]]}}
 
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'''* PcTF transfection for U2OS, outgrowth #1'''
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'''PcTF transfection for U2OS, outgrowth #1'''
 
* 6-well plates: 1-3 = +PcTF, 4-6 = mock (lipo only)
 
* 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.
 
* Expand cells from 6-well plates in individual 10 cm plates (10 mL medium). 6 10 cm plates total.

Latest revision as of 21:45, 26 September 2017

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


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.