Eccles:Antisense Transfection

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Antisense Transfection:

  • For one 24-well:
    • Combine 36μL serum-free media and 4μL Lipofectin in Eppendorf.
    • Incubate for 30 minutes at room temperature.
    • Combine 30μL serum-free media and 10μL 20μM sense or antisense oligo in another Eppendorf.
    • Incubate for 5 - 10 minutes at room temperature.
    • Add Lipofectin dilution to sense/antisense dilution.
    • Incubate for 15 minutes at room temperature.
    • Add 120μL serum-free media.
    • Remove media from well and add 200μL of the above mixture to each well.
    • After 4 hours, add serum to give a final concentration of 10% per well.
    • Culture for the required amount of time.
  • For a 96-well, divide the above volumes by 5, for a 6-well, multiply the above volumes by 5.