User:Karmella Haynes/Notebook/PcTF Genomics/2012/12/03

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12/03/12

  • Cell culture tasks: stock cells and Carly's transfection

U2OS

  • Froze down 5 vials of U2OS (10% DMSO in Tet-free FBS), using 100% confluent 75 cm flask
    • Note: Had Rene transfer from -80°C to -150°C on Thursday
  • Seeded 1:10 and 1:20 75 cm flasks from the small back-up flask


SK-N-SH

  • Froze down 4 vials of SK-N-SH (10% DMSO in Tet-free FBS), using 90% confluent 75 cm flask
    • Note: Had Rene transfer from -80°C to -150°C on Thursday
  • Seeded 1:5 75 cm flask from 1/5 of the original 75 cm flask
  • These cells don't seem to be very robust


K562

  • Seeded 1:10 75 cm flask (passage 5)


K562, Carly's transfection

  • Carly observed RFP signal 24 hours after transfection for all three DNA+ samples; no signal in the three negative controls
  • Pelleted (individually) each of the 6 samples (from 6-well dish) at 1000 rpm/ 3 min.
  • Resuspended in growth medium + pen/strep
  • Plated in a total of 10 mL medium + pen/strep in 10 cm petri dishes



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