Difference between revisions of "Thawing 293FT cells"
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| Line 1: | Line 1: | ||
→ thaw the cells in prewarmed , complete medium without Geneticin | → thaw the cells in prewarmed , complete medium without Geneticin | ||
| − | == Materials | + | == Materials == |
| − | + | * a vial of frozen cells from the liquid nitrogen | |
| − | + | * 37ºC waterbath | |
| − | + | * 75 cm2 flask | |
| − | + | * HS for 293 FT cells | |
| − | + | * 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 09:13, 25 March 2015
→ 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