User:Karmella Haynes/Notebook/PcTF Genomics/2014/09/30

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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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