Bis-Tris Buffer Preparation
Updated 6/12/2015 07:13 PM
Introduction
Materials
- Consumables
- bis-tris
- HCl
- Disposables
- weigh dishes/paper
- pipette tips
- Equipment
- balance
- pH meter
- sterile container
Procedure
Components
Table1
A A | B B | |
1 1 | Final Volume (mL)* | 100 |
2 2 | Final Concentration (M)* | 1 |
Table2
A A | B B | |
1 1 | Component | Amount (g) |
2 2 | bis-tris methane | 20.924 |
3 3 | bis-tris propane | 28.233 |
CRITICALAdd either bis-tris methane or bis-tris propane, only one is required for the buffer.
Preparation
- Add ~70% of the final volume of dH2O to a sterile container.
Generally, the capacity of the container should be greater than (not equal to) the final volume of solution to be prepared in order to prevent the solution from boiling over during the autoclave step.
Ensure you have a volumetric flask measuring your desired final volume.
- Adjust the final volume and molar concentration in the table above and add the calculated amounts to the dH2O.
- Titrate solution to desired pH using HCl.
- Transfer the solution to a sterile volumetric flask and adjust the final volume using dH2O, then return it to the container.
Ensure the solution is well mixed before returning it to the container.
- Apply autoclave tape to the container and autoclave on liquids cycle.
- Store at room temperature.
This protocol describes the preparation of a bis-tris buffer solution.
bis-tris methane buffers between 5.8 ≤ pH ≤ 7.3
bis-tris propane buffers between 6.0 ≤ pH ≤ 9.5