How to Make TE Buffer

We use TE for a number of things in the lab. We dilute and preserve DNA and primers in TE without altering the product. We make our own TE using Tris-HCl and EDTA. We use C1V1=C2V2 along with Molarity formula (M=n/V). Both Tris and EDTA come in powder form and need to be made into a solution. Usually we make a 0.5M EDTA solution and a 1M Tris-HCl solution.

0.5M EDTA (pH 8.0) Materials:

EDTA, MW = 372.24 g/mol 50mL of Autoclaved ddH2O NaOH pellets

Procedure:

1. Use the Molarity formula to find moles 2. Find mass of EDTA using moles and MW 3. Weigh out the mass of EDTA into a beaker 4. Add 40 mL of autoclaved ddH2O and begin mixing. 5. EDTA will only fully dissolve when pH=8. Add NaOH pellets (approximately 10g) to adjust pH to this value. 6. Then top up to 50 mL with ddH2O.

1.0M Tris-HCl (pH 8.0) Materials:

Tris MW = 121.1 g/mol 50mL autoclaved ddH2O HCl

Procedure:
1. 2. 3. 4. 5. 6.

Use the Molarity formula to find moles Find mass of Tris using moles and MW Weigh out the mass of Tris into a beaker Add 40 mL of autoclaved ddH2O and begin mixing. Add HCl with a dropper until pH of 8 is reached Then top up to 50 mL with ddH2O.

TE Buffer (10mM Tris/0.1mM EDTA) Materials:

0.5M EDTA (pH=8.0) 1.0M Tris-HCl (pH=8.0) 50mL Autoclave ddH2O

Procedure:
1. Calculate amount of 0.5M EDTA to add using C1V1=C2V2 (remember 1000mM in 2. 3.
1M) Calculate amount of 1M Tris-HCL to add using C1V1=C2V2 Add appropriate amount of autoclaved ddH2O.

4. Invert bottle 2-3X to mix, then aliquot into 15 mL Falcon tubes and store in the freezer.