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Culturing HEK 293 Cells
Reagents
Medium:
500 ml Dulbeccos Modified Eagle Medium (Gibco #41966-029)
55 ml FCS (10 %)
2.8 ml Gentamycin Solution (Sigma G-1272, 10 ml))
TrypsinTrypsin (10x) lyophilised (Gibco 25095-019)
SolutionVersene 1:5000 (Gibco 15040-033, 100 ml)
Take 2 bottles (2X100 ml) of Versene and pipet 10 ml of each bottle into the bottle with the lyophilised Trypsin to redisolve it. Then redistribute 10 ml of the Trypsin solution back into each Versene bottle.
PBS
Procedures
Starting from a Kryotube:
Prewarm the medium at 37°C and fill two culture flasks (25 ml for 150 cm
Splitting Cells:
Aspirate the medium from the flask. Wash the cells carefully with PBS to remove residual medium. Add 1-2ml of Trypsin Solution (equilibrated at RT) to the flask (150 cm