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Transfection (nonviral)
From 2008.igem.org
Transfection Protocol
Take media off of cells
Rinse with PBS
Put 1mL trypsin on cells. Trypsin will interfere with “sticky” extracellular proteins. When cells start to detach from gelatin, add 5mL media with serum in it. The serum in the media will stop the action of the trypsin.
In multiple-well plate:
Add 1mL media to each well
Count cells with henocytometer to determine concentration of cells.
Add DNA to wells.
Add blasticidin to wells at a one in 1000 volume factor.
Add 105 cells to each well. Shake up and down, side to side
Add 3ul Superfect to each well.
