0
What is a buffer solution?
Buffer solutions are liquids that have both a weak acid and its conjugate base. Due to their composition, the pH (acidity) of the solutions can retain an almost constant degree even though chemical changes occur in solutions. Buffer systems happen in nature, but also highly beneficial in chemistry.
Why we use Buffer Solutions?
Natural buffer solutions maintain a constant pH in organic processes, allowing metabolic processes to take place without hurting the organism. When biologists research biological processes, they need to keep the pH constant, and they use buffer solutions to do so. Biological buffers must fulfill a certain requirement to be useful. They should be soluble in water but not in an organic solvent. They should be unable to cross cell membranes. They must therefore be non-toxic, harmless, and stable in all research for which they are used in.
Buffer solutions are also used in:
- chemical analysis
- dying fabrics
- Fermentation processes
- DNA extraction
- calibration of pH meters
What Is Tris Buffer Solution?
Tris(hydroxymethyl) aminomethane is a chemical compound that is often added to saline because it is non-toxic and is isotonic. Tris buffer solutions, having pKa of 8.1 and a pH range of 7 to 9, are widely used in a variety of biochemical tests and techniques, including DNA extraction. It's necessary to understand that the pH of a tris buffer solution changes as the temperature of the solution increases.
Preparation of Tris Buffer
Tris buffer is easily accessible but if you want to prepare a tris buffer solution, is simple to make with the proper materials and equipment.
Materials:
Calculate the quantity of each product you'll need depending on the molar concentration of the solution and how much buffer you'll need.
- Tris(hydroxymethyl) aminomethane
- HCl
- Distilled deionized water
Procedure:
- To make Tris buffer, first, find out what concentration (molarity) and volume you like. Tris buffer solution for saline, for example, can range from 10 to 100 mM. After you've determined what you're going to make, multiply the molar concentration of the buffer by the volume of the buffer you're going to make to find out how many moles of Tris you'll need.
Molar of Tris=mol/L x L - Then multiply the number of moles by the molecular weight of Tris (121.14 g/mol) to figure out how many grams of Tris there are.
Grams of Tris = Moles x 121.14 g/mol - Dissolves the Tris in 1/3 to 1/2 of your target final volume of distilled deionized water.
- Mix solution in HCl (e.g., 1M HCl) until Tris buffer solution reaches the desired pH.
- Add the water to the buffer to get the desired final volume of solution.
- After preparation, the solution can be stored in a sterile area at room temperature for months. It has a long shelf life because the solution does not contain any proteins.
Uses of Tris Buffer
- Protein Electrophoresis and Western Blotting
Tris buffers are essential to western blotting and protein electrophoresis. Most SDS-PAGE gels, blotting buffers, and running buffers are buffered with Tris.
- Nucleic Acid Agarose Electrophoresis
For DNA agarose electrophoresis, Tris buffers are commonly used. TBE (Tris borate/EDTA) and TAE (Tris-acetate/EDTA) are the two primary buffers. Although there are some variations in the resolution and mobility of different kinds of DNA during electrophoresis, these Tris buffers can be used interchangeably.