Millipore Technical Publications |
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5x Universal Buffer Protocol | ||
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100 mM KCl, 30 mM HEPES pH 7.5, 1 mM MgCl2
Materials needed for 100 mL 5X Universal Buffer:
100 mL sterile container
15 mL conical tube
100 mL RNase-free water
2.24 g Potassium Chloride
0.72 g HEPES-free acid
0.02 g Magnesium Chloride 6H2O
1.20 g Potassium Hydroxide pellets
Protocol:
- Combine 30 mL RNase-free water, 2.24g Potassium Chloride, 0.72 g HEPES-free acid, and 0.02 g Magnesium Chloride 6H2O in a sterile 100 mL container.
- Prepare fresh 2 M KOH by combining 1.20 g Potassium Hydroxide pellets with 10 mL of RNase-free water in a 15 mL conical tube.
- Add 2 mL of 2 M KOH to the solution prepared in step 1.
- Add sterile RNase-free water to a total volume of 100 mL to the solution from step 3 and mix.
- Test pH on a 1 ml aliquot. pH should be in the range of 7.3-7.6.
- Sterile filter or autoclave the solution and store at 4°C for up to 3 months.