Contributor: Suprya Jayadev Date: December 13, 1994 1) Transfect cells. 2) Culture cells 1-3 passages in a T-75 flask containing selection material (e.g. hygromycin, neomycin, etc) 3) Pellet cells and resuspend at a concentration of 1 x 105 cells/ml in selection media. 4) Aliquot 1 drop/well of cell suspension in to each well of a 96-welltissue cultureplate. --> NOTE: Maintain 2-10 plates for each transfection. 5) Check plates every day, if a population grows in a well then subculture cells by diluting 1:10 with selection media and seeding as shown:
6) Continue subculturin in this manner for 5-10 passages to insure that a clonal population is established.
