Tris base
To prepare a 1 M solution, dissolve 121.1 g of Tris base in 800 mL of H2O. Adjust the pH to the desired value by adding concentrated HCl.
| pH | HCl |
|---|---|
| 7.4 | 70 mL |
| 7.6 | 60 mL |
| 8.0 | 42 mL |
Allow the solution to cool to room temperature before making final adjustments to the pH. Adjust the volume of the solution to 1 L with H2O. Dispense into aliquots and sterilize by autoclaving.
If the 1 M solution has a yellow color, discard it and obtain Tris of better quality. The pH of Tris solutions is temperature-dependent and decreases ~0.03 pH units for each 1°C increase in temperature. For example, a 0.05 M solution has pH values of 9.5, 8.9, and 8.6 at 5°C, 25°C, and 37°C, respectively.
[…] To prepare 1 liter of SM buffer, dissolve the NaCl and MgSO4•7H2O in 800 ml of H2O; add the Tris-Cl, and adjust the volume to 1 liter with H2O. Sterilize the buffer by autoclaving for 20 minutes at 15 psi (1.05 kg/cm2) on the liquid cycle. After the solution has cooled, dispense 50-ml aliquots into sterile containers. SM buffer may be stored indefinitely at room temperature. Discard each aliquot after use to minimize the chance of contamination. SM buffer may also be prepared with 0.002% (w/v) gelatin; see the recipe for SM buffer with gelatin. To prepare SM Buffer with gelatin click here and to prepare Tris-cl click here […]
[…] To prepare SM Buffer without gelatin click here and to prepare Tris-cl click here […]