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Selection of Transfected Suspension Cells

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Selection of Transfected Suspension Cells

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-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.