User:Karmella Haynes/Notebook/PcTF Genomics/2014/09/30: Difference between revisions

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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>}}
|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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* Added 80 μL cells to each tube
* Added 80 μL cells to each tube
* LaBaer lab
* LaBaer lab
** Bihazard set-up: Wescodyne waste and rinse; 10 mL in 2x 50 mL conicals
** Biohazard set-up: Wescodyne waste and rinse; 10 mL in 2x 50 mL conicals
** Added 5.0 μL 2.4 mg/mL polybrene (thawed) to each cell sample. Swirled to mix.
** Added 5.0 μL 2.4 mg/mL polybrene (thawed) to each cell sample. Swirled to mix.
** Added 400 μL virus (thawed/ spun/ pipetted to remix) to each cell sample. Swirled to mix. (rinsed pipette tip before discarding)
** Added 400 μL virus (thawed/ spun/ pipetted to remix) to each cell sample. Swirled to mix. (rinsed pipette tip before discarding)
***Note: had some left-over virus; marked these with black dot on caps, placed back into the -80°C box
***Note: had some left-over virus; marked these with black dot on caps, placed back into the -80°C box
** Spun at 2250 rpmn (1178 rcf) for 25 min.
** Spun at 2250 rpm (1178 rcf) for 25 min.
** Pipetted u/d to resuspend, transferred cells to 6 mL plate
** Pipetted u/d to resuspend, transferred cells to 6 mL plate
** Grew at 37°C. Will check GFP expression tomorrow.
** Grew at 37°C. Will check GFP expression tomorrow.

Latest revision as of 00:21, 27 September 2017

Pc-TF Genomics Main project page
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09/30/14

  • K562 transduction (trial 2)
  • Cell culture: passaged U2OS 1:10, K562 1:10
  • Cell culture: started Luc14 & Gal4EED/luc cultures (4 mL 20% FBS/ 1% P/S)

K562 transduction (trial 2)

  • See ##/##/## for transgene/virus list
  • Cell stock passaged 1:10 on 9/22/14. Assume 1.0E6 cells/mL
  • Added 1 mL complete culture medium to 6x 15 mL conicals
  • Added 80 μL cells to each tube
  • LaBaer lab
    • Biohazard set-up: Wescodyne waste and rinse; 10 mL in 2x 50 mL conicals
    • Added 5.0 μL 2.4 mg/mL polybrene (thawed) to each cell sample. Swirled to mix.
    • Added 400 μL virus (thawed/ spun/ pipetted to remix) to each cell sample. Swirled to mix. (rinsed pipette tip before discarding)
      • Note: had some left-over virus; marked these with black dot on caps, placed back into the -80°C box
    • Spun at 2250 rpm (1178 rcf) for 25 min.
    • Pipetted u/d to resuspend, transferred cells to 6 mL plate
    • Grew at 37°C. Will check GFP expression tomorrow.




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