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Tris buffer is one of the most widely used buffering agents in biochemical research, and its common effective pH range is in the "neutral" range, such as:
Tris buffer: pH=7.5~9
Tris-HCl buffer: pH = 7.5~8.5
Tris-phosphate buffer: pH = 5.0~9.0
1. Preparation of 0.05mol/L Tris buffer for a specific pH value: Mix 50ml of 0.1mol/L Tris base solution with the corresponding volume (unit: ml) of 0.1ml HCl as shown in the table below, and add water to adjust the volume to 100ml.
2. Two configuration methods of Tris-HCl buffer
The first method is to prepare 0.05 mol/L Tris and 0.05 mol/L HCl solutions, respectively, and then mix them according to the volume listed in the common table.
However, as the standard concentration of dilute hydrochloric acid is not easy to prepare, another method is commonly used: take the configuration of 1 L of 0.1 mol/L Tris-HCl buffer as an example: Firstly, 12.11 g Tris base was dissolved in 950 mL ~ 970 mL deionized water, and 4 N HCl was added by drop while stirring. The pH value of the solution was measured by pH meter to the desired pH value, and then water was added to make up to 1 L.
3. Other TRIS-derived buffers
In addition to Tris-HCl, there are a variety of TRIS-derived buffers:
TBS=Tris-HCl+ NaCl+KCl, which is often used to clean immunostained tissues or Western blot membranes in Western Blotting.
TBST=Tris-HCl+NaCl+tween20, a kind of membrane washing buffer commonly used in Western Blotting;
TE=Tris-HCl+EDTA, which protects the bases of DNA and is often used for DNA stabilization and storage;
TAE=Tris base+ acetic acid +EDTA is a widely used buffer system for short time electrophoresis of large DNA fragments.
TBE=Trisbase+ boric acid +EDTA, suitable for long time DNA electrophoresis, small fragment separation effect is better.
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