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This protocol has not been updated this year! Last update was on December 11, 2025.
DNase (2X) and papain (2X)
DNase and papain from 2X
This is based off of Worthington Papain Dissociation System to achieve a final ** 20 U papain/mL + 2,000 U DNase I/mL solution with 0.5mM EDTA and 1mM L-cysteine**.
Add equal volumes 2X DNase and papain from the -80°C. e.g. 1 mL DNase (2X) + 1 mL papain (2x).
Solution |
Details |
|---|---|
4,000 U/mL in DMEM/F12 |
|
40 U/mL with 1mM EDTA, 2mM L-cysteine in DMEM/F12 |
L-Cysteine (50 mM)
Measure 60.6 mg L-Cysteine (MW: 121.16 g/mol) and add to a 15 mL conical
Add 10 mL DMEM/F12
Take into TC and sterilize by filtering through a 0.22 µm filter
Aliquot ~1.1 mL into 1.6 mL eppendorf tubes and freeze with papain aliquots at -80°C
Papain (2X: 1mM EDTA, 2mM L-cysteine, 40 U papain/mL)
To make a 50 mL solution of 2X papain from Papain 100 mg:
Component |
Final conc |
Volume |
|---|---|---|
EDTA (0.5M, pH=8.0) |
1 mM |
100 µL |
L-Cysteine (50mM) |
2 mM |
2 mL |
Papain (100 mg bottle) |
40 U/mL |
100 µL |
DMEM/F12 |
47.9 mL |
Sterilize with 0.22 µM filter and store aliquots at -80°C. EDTA (0.5M, pH=8.0) should be in Bruni and L-Cystein (50mM) should be in -80°C with papain aliquots.
DNase (2X: 4,000 U/mL solution)
To make a 2X solution from DNase I (100 mg, 2000 units/mg).
Add 50 mL DMEM/F12 to a 100 mg bottle, sterilize with 0.22 µM filter and store aliquots at -80°C.