How to Prepare a 250 ml Solution of 50 mM Tris at pH 8.0

To prepare a 250 ml solution of 50 mM Tris buffer at pH 8.0, follow these steps:

1. Calculate the Amount of Tris Needed:
   To make a 50 mM solution, you will need:

50 mM is equivalent to 0.050 g of Tris per liter. For 250 ml, you calculate:

0.050 g/L × 0.250 L = 0.0125 g

So, you need to weigh out 0.0125 g of Tris powder.

2. Dissolve the Tris in Water:
   Add 0.0125 g of Tris to about 100 ml of high-purity water in a clean container. Stir well to make sure the powder fully dissolves.
3. Adjust the pH:
   Using a pH meter, carefully adjust the pH of the solution to 8.0. To do this, add 0.1 M HCl dropwise while mixing until the desired pH is achieved. Make sure to measure the pH continuously during this step.
4. Complete the Volume:
   Once the pH is adjusted, transfer the solution into a 250 ml volumetric flask or measuring cylinder. Add high-purity water until the total volume reaches 250 ml. Make sure to mix thoroughly after reaching the final volume.

Now you have a 250 ml solution of 50 mM Tris at pH 8.0, ready for your experiments!