Preparation of High Serum for A1 cells: Difference between revisions
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(Created page with "For 400 ml FCS 40 ml Eagle’s MEM 252 ml DdH2O 100 ml Pen/ Strep 4 ml Glutamin 4 ml Insulin (stock: 1mg/ ml) 4 ml - grow at 25ºC in 2% CO2 - if you want to gr...") |
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For 400 ml | [[For 400 ml]] | ||
FCS 40 ml | FCS 40 ml | ||
Revision as of 10:09, 13 February 2015
FCS 40 ml Eagle’s MEM 252 ml DdH2O 100 ml Pen/ Strep 4 ml Glutamin 4 ml Insulin (stock: 1mg/ ml) 4 ml
- grow at 25ºC in 2% CO2 - if you want to grow without CO2, then substitute L-15 for the MEM
- passage once per week 1:3
- wash with APBS (100 ml PBS + 25 ml ddH2O) - trypsinize with TE - stop with media - centrifuge cells at 100 rpm 3 minutes
to make myotubes:
- wash cells 1x with APBS - put cells in 0.5% FCSmedia - wait 3 days