LiberaseTM cell dissociation for axolotl/frog limb cells
Prepare Reagents
0.7x PBS
- Dilute 1x PBS to 0.7x with dH2O
- -> Chill on ice before use.
Liberase TM solution
- 40uL Stock Liberase TM (100x, 26WU/mL)
- 3960uL 0.7xPBS
- -> Chill on ice before use.
AMEM(0) (serum-free medium)
- 114.3mL F12:DMEM+Glutmax
- 1.5mL Sodium pyruvate (100mM)
- 1.5mL B27 supplement
- 1.5mL Insulin (1 mg/mL)
- 1.5mL MEM Non-Essential Amino Acid (NEAA)
- 1.5mL Penicillin/Streptomycin (10000 U/mL)
- 28.2mL Sterile ddH2O
- -> Total 150mL, filter.
- -> Chill on ice before use.
High-Serum AMEM (HS-AMEM)
- 125mL Minimum essential medium (MEM)
- 20mL Heat-inactivated fetal bovine serum (56 degree, 30 min)
- 2mL Insulin (1 mg/mL)
- 2mL Glutamine (200mM)
- 2mL Penicillin/Streptomycin (10000 U/mL)
- 49mL Sterile ddH2O
- -> Total 200mL, filter.
- -> Chill on ice before use.