How to Prepare Tris EDTA (TE) Buffer?

TE buffer (Tris-EDTA) is a typical buffer solution for the resuspension and storage of nucleic acids, particularly DNA. The Tris solution maintains the DNA soluble in water, while EDTA, a cation chelator, protects nucleic acids from enzymatic destruction.

Reagents For the Preparation of TE (Tris EDTA) Buffer

• 1M Tris-HCl (pH 8)
• 0.5M EDTA (pH 8)
• Distilled water

Equipment Required For the Preparation of TE Buffer

• Autoclave
• Pipette
• Pipette tips
• Duran bottle

Procedure For the Preparation of TE Buffer

• Step 1: 1mL of 1M Tris-Cl (pH 8.0) should be added to one 100-ml Duran bottle.

• Step 2: Measure out 0.2 ml of 0.5M EDTA (pH 8.0) and add it to the Duran bottle.

• Step 3: With 98.8 ml of distilled water, top up the solution to 100 ml.

• Step 4: After covering the bottle with the lid, shake it several times to combine swiftly.

• Step 5: Autoclave the solution on a liquid cycle for 20 minutes at 15 psi to disinfect it.

Preparation of 1x TE (Liquid Reagents)

Combine the following materials in a graduated cylinder or flask:

• 10ml of 1M Tris-HCl at the final desired pH
• 2ml of 0.5M EDTA pH8
• 988ml of distilled or deionized water

Procedure For Preparation of 1x TE (Liquid Reagents)

• Add a magnetic stir bar and mix the above-mentioned ingredients for 2 minutes on a hot plate with heat set to “off” and stir set to “300 RPM”.
• In a fume hood, check pH and make any required adjustments as per the instructions. 
• An autoclave or 0.22µm filter can be used to sterilize the fluid.

Preparation of 10x TE (Liquid Reagents)

Combine the following materials in a graduated cylinder or flask:

• 100ml of 1M Tris-HCl at the final desired pH
• 20ml of 0.5M EDTA pH8
• 880ml of distilled or deionized water

Procedure For Preparation of 10x TE (Liquid Reagents)

• Add a magnetic stir bar and mix the above-mentioned ingredients for 2 minutes on a hot plate with heat set to “off” and stir set to “300 RPM”.
• In a fume hood, check pH and make any required adjustments as per the instructions. 
• An autoclave or 0.22µm filter can be used to sterilize the fluid.

Preparation of 1x TE (Dry Reagents, Tris-HCl)

Combine the following materials in a graduated cylinder or flask:

• 1.211g of Tris Base (MW 121.14)
• 0.292g of EDTA (MW 292.25)
• 800ml of distilled or deionized water

Procedure For Preparation of 1x TE (Dry Reagents, Tris-HCl)

• Add a magnetic stir bar and mix the above-mentioned ingredients for 5 minutes on a hot plate with heat set to “off” and stir set to “300 RPM”.
• In a fume hood, check pH and make any required adjustments as per the instructions. 
• The solution can be sterilized using an autoclave or a 0.22µm filter.

Preparation of 1x TE (Dry Reagents, Tris-Base)

Combine the following materials in a graduated cylinder or flask:

• 12.11g of Tris Base (MW 121.14)
• 2.92g EDTA (MW 292.25)
• 800ml of distilled or deionized water

Procedure For Preparation of 1x TE (Dry Reagents, Tris-Base)

• Add a magnetic stir bar and mix the above-mentioned ingredients for 5 minutes on a hot plate with heat set to “off” and stir set to “300 RPM”.
• In a fume hood, check pH and make any required adjustments as per the instructions. 
• The solution can be sterilized using an autoclave or a 0.22µm filter.

Storage of TE Buffer

TE buffer should be stored at room temperature (15 °C to 25 °C).

Safety Hazards

The TE buffer is not considered potentially hazardous. However, before using, please read the TE buffer safety data sheet.

Video References

References

• https://www.aatbio.com/resources/buffer-preparations-and-recipes/te-buffer-10x
• https://toptipbio.com/te-buffer-recipe/
• https://cshprotocols.cshlp.org/content/2009/2/pdb.rec11661.short
• https://static.igem.org/mediawiki/2021/3/36/T–Thessaloniki–Preparation_of_TE_Buffer.pdf
• https://gatescientific.com/technique-geeks-blog/f/recipes-for-te-tris-edta