LiberaseTM cell dissociation for axolotl/frog limb cells: Difference between revisions
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(Created page with "= Prepare Reagents = ---- ‘’’0.7x PBS’’’ *Dilute 1x PBS to 0.7x with dH2O :-> Chill on ice before use") |
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= Prepare Reagents = | = Prepare Reagents = | ||
== 0.7x PBS == | |||
*Dilute 1x PBS to 0.7x with dH2O | *Dilute 1x PBS to 0.7x with dH2O | ||
:-> Chill on ice before use | :-> 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. | |||
Revision as of 21:48, 18 July 2022
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.