Thawing 293FT cells: Difference between revisions
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a vial of frozen cells from the liquid nitrogen
37ºC waterbath
75 cm2 flask
HS for 293 FT cells
pipetts
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→ thaw the cells in prewarmed , complete medium without Geneticin | → thaw the cells in prewarmed , complete medium without Geneticin | ||
materials: | == materials: == | ||
<li>a vial of frozen cells from the liquid nitrogen | |||
<li>37ºC waterbath | |||
<li>75 cm2 flask | |||
<li>HS for 293 FT cells | |||
<li>pipetts | |||
procedure: | == procedure: == | ||
# remove the vial of frozen cells from liquid nitrogen and thaw quickly in a 37ºC waterbath | # 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 | # transfer the cells to a 75 cm2 flask containing 12 ml Hs for 293 FT cells w/o Geneticin | ||
Revision as of 08:02, 22 August 2014
→ thaw the cells in prewarmed , complete medium without Geneticin
materials:
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