0.1 M Sodium Phosphate Buffer Preparation
Introduction
Sodium phosphate buffer is a common buffer solution used in various biochemical and molecular biology applications. It is a mixture of monobasic and dibasic sodium phosphate salts, which provides a buffering capacity to maintain a stable pH. In this article, we will describe the preparation of 0.1 M sodium phosphate buffer solution.
Materials
- Sodium phosphate monobasic (NaH2PO4): 13.8 g
- Sodium phosphate dibasic (Na2HPO4): 26.8 g
- Distilled water: 1 L
Protocol
Step 1: Preparation of 0.1 M Sodium Phosphate Monobasic Solution
- Weigh 13.8 g of sodium phosphate monobasic (NaH2PO4) and transfer it to a 1 L beaker.
- Add approximately 500 mL of distilled water to the beaker and stir until the salt is completely dissolved.
- Bring the volume to 1 L by adding more distilled water.
Step 2: Preparation of 0.1 M Sodium Phosphate Dibasic Solution
- Weigh 26.8 g of sodium phosphate dibasic (Na2HPO4) and transfer it to a separate 1 L beaker.
- Add approximately 500 mL of distilled water to the beaker and stir until the salt is completely dissolved.
- Bring the volume to 1 L by adding more distilled water.
Step 3: Preparation of 0.1 M Sodium Phosphate Buffer Solution
- Mix 50 mL of 0.1 M sodium phosphate monobasic solution with 50 mL of 0.1 M sodium phosphate dibasic solution.
- Stir well to combine the two solutions.
- Adjust the pH of the buffer solution to the desired value (e.g., pH 7.0) using a pH meter.
Notes
- The preparation of 0.1 M sodium phosphate buffer solution requires careful attention to the pH, as it can affect the activity of enzymes and other biological molecules.
- The buffer solution can be stored at room temperature for up to 3 months.
- It is recommended to filter the buffer solution through a 0.22 μm filter before use to remove any impurities.
Conclusion
In this article, we have described the preparation of 0.1 M sodium phosphate buffer solution. This buffer solution is commonly used in various biochemical and molecular biology applications, including protein purification, PCR, and Western blotting. By following the protocol outlined above, you can prepare a high-quality buffer solution that meets your research needs.
