Haynes Lab:Notebook/Short Projects/2014/09/11

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9/11/14 - Jonah Thomas

  • Cell culture - check cells, add dox to dox+ cells
  • Plan transfections - measure concentration of plasmid DNA

Cell culture

  • Checked cells. Cells are attached, about 50 - 60 % confluent. Will do Transfections on Friday.
  • Added 1 mg/mL doxycycline (dox) to a final concentration of 1 μg/mL to dox+ 6-well plate and T-75 flask
    • Added 4 μL dox directly to wells in 6-well plate
    • Transferred 2 mL medium from flask to 15 mL conical, added 10 μL dox, added 2 mL back to flask with sterile pipette


Plan Transfections


Measurement (Biotek machine - Take 3 plate)

  • OD260 = 0.384
  • 260/280 = 1.88
  • ng/μL = 384.2


Strategy for transfection: add Lipo/DNA to 6 wells total...

  • Will need 2000 ng KAH60/pcVN DNA per transfecttion
  • Plate 1 - no dox, transfect 3 wells only
  • Plate 2 - dox+, transfect 3 wells only



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