Preparation of High Serum for C2C12 cells

Material

• 1 tube pen/strep (green, 4 ml)
• 1 tube glutamine (blue, 4 ml) → this is not necessary if the DMEM has glutamax
• 1 bottle DMEM with 4500 mg/ l glucose
• 40 ml FCS (frozen in a 50 ml FALCON tube)
• 1 large Stericup (500 ml) (bottle + filter + top)

Procedure

1. thaw all frozen stuffs at 37 ºC
2. add in the Stericup the 40 ml FCS, 4 ml pen/ strep and the 4 ml glutamine (if necessary)
3. fill this up to 400 ml with the DMEM
4. filtrate the Medium in the bottle by connecting the vacuum pump to the steritop
5. label the bottle with the name, the date and “high serum for C2C12”