Eccles:Antisense Transfection

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Antisense Transfection:

 * For one 24-well:
 * Combine 36&mu;L serum-free media and 4&mu;L Lipofectin in Eppendorf.
 * Incubate for 30 minutes at room temperature.
 * Combine 30&mu;L serum-free media and 10&mu;L 20&mu;M 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 120&mu;L serum-free media.
 * Remove media from well and add 200&mu;L 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.