TAE buffer Totally Explained

Everything about Tae Buffer totally explained

TAE buffer is a buffer solution used in agarose electrophoresis, typically for the separation of nucleic acids such as DNA and RNA. It is made up of Tris-acetate buffer, usually at pH 8.0, and EDTA, which sequesters divalent cations. TAE has a lower buffer capacity than TBE and easily can become exhausted, but linear, double stranded DNA runs faster in TAE.

Uses

TAE buffer is used as both a running buffer and in agarose gel. TAE has been used at various concentrations to study free DNA solution mobility with and without Sodium Chloride. Use in denaturing gradient gel electrophoresis methods for broad-range mutation analysis has also been described.

Preparation

For 1 litre of 50x TAE buffer use:
- 242 g Tris base (2-amino-2-hydroxymethyl-propane-1,3-diol) (= 2 mole) - 57.1 ml glacial acetic acid (= 100% acetic acid) (57.19 ml = 1 mole) - 100 ml 0.5 M Na₂ EDTA (pH 8.0) - H₂O up to 1000 ml
To prepare 0.5 M Na₂ EDTA (pH 8.0) add 186.1 g of disodium ethylenediaminetetraacetate x 2H₂O to 800 ml of H₂O. Stir vigorously. Adjust the pH to 8.0 with NaOH (ca. 20 g of NaOH). Sterilize by autoclaving. Hint: The disodium salt of EDTA won't go into solution until the pH of the solution is adjusted to ca. 8.0 by the addition of NaOH.

Reference

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