How to Prepare Tris EDTA (TE) Buffer?

How to Prepare Tris EDTA (TE) Buffer
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

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References

About Author

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Jyoti Bashyal

Jyoti Bashyal, a graduate of the Central Department of Chemistry, is an avid explorer of the molecular realm. Fueled by her fascination with chemical reactions and natural compounds, she navigates her field's complexities with precision and passion. Outside the lab, Jyoti is dedicated to making science accessible to all. She aspires to deepen audiences' understanding of the wonders of various scientific subjects and their impact on the world by sharing them with a wide range of readers through her writing.

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