Thawing 293FT cells: Difference between revisions
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(Created page with "→ thaw the cells in prewarmed , complete medium without Geneticin materials: - a vial of frozen cells from the liquid nitrogen - 37ºC waterbath - 75 cm2 flask - HS for 293...") |
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procedure: | procedure: | ||
# remove the vial of frozen cells from liquid nitrogen and thaw quickly in a 37ºC waterbath | |||
# transfer the cells to a 75 cm2 flask containing 12 ml Hs for 293 FT cells w/o Geneticin | |||
# incubate the flask for 2 –4 hours to allow the cells to attach to the bottom of the flask | |||
# aspirate off the medium and replace with 12 ml of fresh, complete medium without Geneticin | |||
# incubate the cells over night | |||
# the next day, aspirate off the medium and replace with fresh, complete medium containing 500 µg/ ml Geneticin | |||
# incubate the cells and check them daily until the cells are 80 – 90% confluent | |||
# proceed to subculturing cells | |||
Revision as of 07:56, 22 August 2014
→ thaw the cells in prewarmed , complete medium without Geneticin
materials: - a vial of frozen cells from the liquid nitrogen - 37ºC waterbath - 75 cm2 flask - HS for 293 FT cells - pipetts
procedure:
- remove the vial of frozen cells from liquid nitrogen and thaw quickly in a 37ºC waterbath
- transfer the cells to a 75 cm2 flask containing 12 ml Hs for 293 FT cells w/o Geneticin
- incubate the flask for 2 –4 hours to allow the cells to attach to the bottom of the flask
- aspirate off the medium and replace with 12 ml of fresh, complete medium without Geneticin
- incubate the cells over night
- the next day, aspirate off the medium and replace with fresh, complete medium containing 500 µg/ ml Geneticin
- incubate the cells and check them daily until the cells are 80 – 90% confluent
- proceed to subculturing cells