9/11/14 - Jonah Thomas
- Cell culture - check cells, add dox to dox+ cells
- Plan transfections - measure concentration of plasmid DNA
- 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
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