Proteinase K Recipe
Proteinase K or ProK is a common, essential reagent for DNA Extraction Protocols. Here we provide a recipe for generating a 20mg/mL ProK solution in 50mM Tris-HCl (pH 8.0), 3mM CaCl2.
Reagents Required:
- Powdered Proteinase K (GoldBio Cat #P-480-1)
- 1M Tris-HCl aq. soln. (Thermo Fisher Cat#J22638.AE)
- 1M CaCl2 aq. soln. (Thermo Fisher Cat#J63122)
| 1mL | 10mL | |
|---|---|---|
| Proteinase K poweder (g) | 0.02g or 20mg | 0.2g or 200mg |
| 1M Tris-HCl (µL) | 50µL | 500µL |
| 1M CaCl2 (µL) | 3µL | 30µL |
Instructions
- Add Tris-HCl and CaCl2 to either a 1.5mL tube or a 10mL tube, depending on volume
- Carefully measure either 0.02g or 0.2g of Proteinase K powder
- Add a small amount of ddH2O (~300µL/3mL) to tube
- Add Proteinase K, in several scoops, vortexing each time to dissolve
- Add enough ddH2O to reach 1mL or 10mL mark, depending on recipe
- Aliquot 1.5mL into brand new 1.5mL tubes, label each with “pro-k, date, tube #”
Store aliquots in -20ºC