HOW TO MAKE TRIS HCL BY DR YOUSIF

1. Dr YOUSIF HAMED MOHAMED
SHARIF
How to make Tris-HCl buffer solution
Tris(hydroxymethyl)aminomethane hydrochloride (Tris-HCl) is a commonly
used buffering solution. The role of Tris-HCl in molecular biology is to control
the acidity and osmolarity of a solution. Due to its properties, Tris-HCl is
often a component of lysis buffers. The pH of the solution is most commonly
made between 7.0 and 8.0.
1 M Tris-HCl recipe
The recipe below is used to prepare a 100 mL 1 M Tris-HCl solution at pH
8.0. It can, however, be tweaked to make the same solution at the desired
pH.
Reagent Weight / Volume Final concentration
Tris base 12.11 grams 1 M
Distilled H2O Up to 100 mL
How to make 1 M Tris-HCl pH 8.0
1. Weigh out 12.11 g Tris and add to a 100 mL Duran bottle.
2. Measure out 80 mL of distilled water and add to the Duran bottle.
3. Add a magnetic flea and place on a magnetic stirring plate to mix
the solution.
4. Add a pH meter into the solution to observe the pH.
5. Slowly add concentrated hydrochloric acid (HCl) solution using a
Pasteur pipette to reduce the pH to 8.0, or another desired pH. Be

2. careful not to add too much at a time, since the pH will change
rapidly.
6. Once the desired pH has been reached, top up the solution to 100
mL using distilled water.
7. To sterilise, autoclave the solution on a liquid cycle (20 min at 15
psi).
Storage of 1 M Tris-HCl pH 8.0 solution
Store 1 M Tris-HCl pH 8.0 solutions at room temperature (+15o
C – +25o
C).
Safety
Tris base is not classed as a hazardous substance, however, be careful when
using HCl during the pH adjustment step. Concentrated HCl solution is an
acid as is very corrosive. Always read the material safety data sheet before
using it.
Reference
https://toptipbio.com/tbe-buffer-10x-recipe/