Altman:Protocols/TC Protocols/Splitting ARPE-19

Department of Physics, Willamette University

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Freezing down SF9 cells for long-term storage

1. Remove the cell media from the cells

2. Wash the cells with 5 mL of PBS

3. Remove PBS, and put in 5 mL of Trypsin

4. Incubate for 10 minutes at 37° and 5% CO2

5. Look at the cells using the microscope; you should see the cells detaching from the surface

6. Knock the flask to dislodge the cells

7. Put in 1 mL of FBS

8. Put the 6 mL of cells into a conical

9. Spin at 500 rpm for 3 minutes

10. Pull off the supernatant (6 mL)

11. Re-suspend in 10 mL of media