Tris-Cl Stock Preparation
Updated 6/12/2015 07:56 PM
Introduction
Materials
- Consumables
- tris-base
- hydrogen chloride (concentrated)
- Disposables
- weigh dishes/paper
- Equipment
- balance
- media bottle
- pH meter
Procedure
Component Composition
Table1
A A | B B | |
1 1 | Final Volume (mL)* | 1000 |
2 2 | Final Concentration (M)* | 1 |
Table3
A A | B B | |
1 1 | Component | Weight (g) |
2 2 | tris-base | 121.14 |
Preparation
- Add ~80% of the final volume of dH2O to a sterile media bottle.
Generally, the capacity of the bottle should be greater than (not equal to) the final volume of media to be prepared in order to prevent the media 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 of tris-base to the dH2O.
Solution likely will not completely dissolve until at correct pH value.
- Titrate solution to desired pH using concentrated HCl.
- Once completely dissolved, transfer the solution to a sterile volumetric flask and adjust the final volume using dH2O, then return it to the media bottle.
Ensure the solution is well mixed before returning it to the media bottle.
- Apply autoclave tape to the bottle and autoclave on liquids cycle.
CRITICALLiquids cycle.
- Store at room temperature.
This protocol is for the preparation of tris(hydroxymethyl)aminomethane (Tris) at various concentrations.
Note: Tris-base solubility in water ~4M (25°C).
Sambrook, J. & Russell, D. W. (2001). Molecular Cloning: A Laboratory Manual, 3 edn. Cold Spring Harbor, N.Y.: Cold Spring Harbor Laboratory Press. pp. A1.7