How to make a buffer 10X Stock Solution of TE Buffer, pH 8.0

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Tris EDTA buffer is commonly used for nucleic acids to neutralize and stabilize the pH. Use to isolate, purify, and store DNA, cDNA, or RNA for cloning, qPCR, or gel electrophoresis. Here’s how to prepare a 10X TE stock solution that can easily be diluted 1:10 to make 1X TE buffer.
1. Weigh out 1.6g EDTA (0.01M) and 7.88g Tris-HCl or Trizma® Hydrochloride (0.1M).
2. Add 400 mL ultrapure Milli-Q ® H2O to a beaker with a stir bar.
3. Add Tris-HCl and EDTA, stirring until fully dissolved.
4. Use a calibrated pH meter to measure and adjust the buffer pH with HCl and NaOH to pH 8.0. Be sure to exercise caution when working with concentrated acids and bases.
5. Pour buffer into a volumetric flask and q.s. to a final volume of 500 mL with Milli-Q ® ultrapure H2O.
6. Mix well.
7. Label and store the 10X TE stock solution at room temperature or 4°C for 3 - 6 months.
8. Dilute 10X TE stock 1:10 to make fresh 1X TE working buffer for your assay. Also, you can autoclave or sterile filtered the buffer according to your lab and assay requirements.
EDTA: www.sigmaaldri...
Trizma® Hydrochloride: www.sigmaaldri...
HCl: www.sigmaaldri...
NaOH: www.sigmaaldri...
Milli-Q® Water Purification Systems: www.sigmaaldri...
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10X Tris EDTA: www.sigmaaldri...

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6 сен 2024

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Комментарии : 12

@Its_InduB Год назад

In TE,tris base is used or tris hcl; i am confused. And why the molarities of tris and edta is different. Please explain.

@MerckLifeScience Год назад

Hi Indu! TE buffer is also known as T10E1 buffer due to the working concentration of Tris and EDTA. Therefore, 10X TE buffer is 0.1M Tris HCl and 0.01M EDTA. Tris or Trizma® base is alkaline and stable with or without HCl. However, Tris or Trizma® base alone is much more alkaline, pH 10 - 12. Whereas Tris-HCl has a buffering capacity of pH 7 - 9 which is much closer to the target for the TE buffer we prepared at pH 8. EDTA also has a pH of 8 and this is where DNA & RNA are stable. Please let us know if you have any other questions!

@Its_InduB Год назад

@@MerckLifeScience thanks for the reply."but why these two should be in different molarities?"

@MerckLifeScience Год назад

Hi Indu - The two components serve distinct purposes in the buffer and therefore have different molarity requirements. TE buffer’s alternate name, T10E1 Buffer, signifies the concentration ratio of its two components, Tris and EDTA, which are present at 0.1M and 0.01M, respectively. Tris serves to maintain the buffer's capacity, while EDTA acts as a chelating agent, effectively binding metal ions and rendering them unavailable for degrading the DNA/RNA. Let us know if you have any other questions!

@nm8734 9 месяцев назад

How you calculate tris hcl

@MerckLifeScience 9 месяцев назад

Thanks for your comment! To prepare 0.1M Tris-HCl, remember, 0.1M is the same as 0.1 mol/L. We are making 0.5L and will need the molecular weight (MW) for Tris-HCl: 157.594 g/mol. Let’s calculate: 0.1 mol/L x 0.5L = 0.05 mol. Next, multiply by MW: 0.05 mol x 157.594 g/mol = 7.88 g Tris-HCl Please let us know if you have any other questions! Happy to help.

@mohanchaitanyareddy2577 Год назад

For how many days does 10X TE can be stored?

@MerckLifeScience Год назад

The 10X TE stock can be stored up to 3 months at room temperature, but can also be refrigerated and used as needed to prepare 1X TE working solution. Please let us know if you have any other questions!

@radosny762 10 месяцев назад

Autoclave it?

@MerckLifeScience 10 месяцев назад

Thank you for your question! Yes, it can be autoclaved or sterile filtered if desired, depending on your application and standard lab practices. After autoclaving, remember to let it cool and check the pH to ensure it hasn't changed.

@jayalakshmi5306 Год назад

To store dna should I use 10x or 1x TE buffer

@MerckLifeScience Год назад

It's not recommended to resuspend DNA in 10X as it's too concentrated with salt. Instead, dilute the buffer stock 1:10 for a 1X TE working solution for resuspension or dilution of purified DNA or RNA. For storage, you should freeze it at -20 to -70C, but avoid too many freeze thaws. One tip is to make several aliquots for your different assays. Please let us know if you have any other questions!