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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→ thaw the cells in prewarmed , complete medium without Geneticin
→ thaw the cells in prewarmed , complete medium without Geneticin


materials:
== Material ==
- a vial of frozen cells from the liquid nitrogen
* a vial of frozen cells from the liquid nitrogen
- 37ºC waterbath
* 37ºC waterbath
- 75 cm2 flask
* 75 cm2 flask
- HS for 293 FT cells
* HS for 293 FT cells
- pipetts
* pipetts


procedure:
== Procedure ==
 
# remove the vial of frozen cells from liquid nitrogen and thaw quickly in a 37ºC waterbath
1) 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
2) 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
3) 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
4) aspirate off the medium and replace with 12 ml of fresh, complete medium without  Geneticin
# incubate the cells over night
5) incubate the cells over night
# the next day, aspirate off the medium and replace with fresh, complete medium containing 500 µg/ ml Geneticin
6) 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
7) incubate the cells and check them daily until the cells are 80 – 90% confluent
# proceed to subculturing cells
8) proceed to subculturing cells

Latest revision as of 14:18, 8 June 2015

→ thaw the cells in prewarmed , complete medium without Geneticin

Material

  • a vial of frozen cells from the liquid nitrogen
  • 37ºC waterbath
  • 75 cm2 flask
  • HS for 293 FT cells
  • pipetts

Procedure

  1. remove the vial of frozen cells from liquid nitrogen and thaw quickly in a 37ºC waterbath
  2. transfer the cells to a 75 cm2 flask containing 12 ml Hs for 293 FT cells w/o Geneticin
  3. incubate the flask for 2 –4 hours to allow the cells to attach to the bottom of the flask
  4. aspirate off the medium and replace with 12 ml of fresh, complete medium without Geneticin
  5. incubate the cells over night
  6. the next day, aspirate off the medium and replace with fresh, complete medium containing 500 µg/ ml Geneticin
  7. incubate the cells and check them daily until the cells are 80 – 90% confluent
  8. proceed to subculturing cells
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