Contributor: Suprya JayadevDate: December 13, 19941) 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-well tissue culture plate.--> 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.