How to Prepare 1x TE Buffer
The Tris-EDTA (TE) buffer is a commonly used solution in molecular biology and biochemistry laboratories. It is primarily used to maintain the stability of nucleic acids and to facilitate the separation of DNA and RNA during electrophoresis. Preparing a 1x TE buffer is a straightforward process that requires a few basic laboratory supplies and reagents. In this article, we will guide you through the steps to prepare a 1x TE buffer.
Materials Needed
Before you begin, make sure you have the following materials on hand:
– Distilled water
– Tris base (Tris hydroxymethyl aminomethane)
– EDTA (ethylene diamine tetraacetic acid) disodium salt
– Acetic acid
– pH meter
– Pipettes and tips
– Beakers or tubes
– Safety equipment (gloves, lab coat, etc.)
Step-by-Step Instructions
1. Calculate the volumes of Tris base and EDTA needed: To prepare a 1x TE buffer, you will need 10 mM Tris-HCl and 1 mM EDTA. For a 1-liter solution, you will need 1.21 g of Tris base and 37.2 mg of EDTA.
2. Weigh the Tris base and EDTA: Using a balance, weigh out the required amounts of Tris base and EDTA. Be sure to handle these chemicals with care, wearing gloves and a lab coat.
3. Dissolve the Tris base in distilled water: Add the Tris base to a beaker or tube containing 100 mL of distilled water. Stir the mixture until the Tris base is completely dissolved.
4. Dissolve the EDTA in distilled water: In a separate beaker or tube, dissolve the EDTA in 50 mL of distilled water. Stir the mixture until the EDTA is completely dissolved.
5. Combine the solutions: Carefully pour the EDTA solution into the Tris solution. Stir the mixture thoroughly to ensure that the EDTA is evenly distributed.
6. Adjust the pH: Use acetic acid to adjust the pH of the TE buffer to 8.0. A pH meter will be helpful in this step to ensure the correct pH is achieved.
7. Dilute the solution to 1 liter: Once the pH is adjusted, add distilled water to the solution until the total volume reaches 1 liter. Stir the mixture to ensure that the buffer is well-mixed.
8. Filter the buffer: It is recommended to filter the TE buffer through a 0.22 µm filter to remove any particulates that may be present.
9. Store the buffer: Store the prepared TE buffer in a labeled container at room temperature. TE buffer can be stored for several months if properly stored.
By following these steps, you will have successfully prepared a 1x TE buffer that can be used for various molecular biology applications. Always remember to practice proper laboratory safety and follow the guidelines for handling chemicals.
