TAE buffer

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TAE buffer is a buffer solution used in agarose electrophoresis, typically for the separation of nucleic acids such as DNA and RNA.[1] 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.

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[edit] Uses

TAE buffer is used as both a running buffer and in agarose gel.[2] TAE has been used at various concentrations to study free DNA solution mobility with and without Sodium Chloride.[3] Use in denaturing gradient gel electrophoresis methods for broad-range mutation analysis has also been described.[4]

[edit] 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 will not go into solution until the pH of the solution is adjusted to ca. 8.0 by the addition of NaOH.

[edit] Reference

  1. ^ Ogden, R.C., and Adams, D.A., Electrophoresis in agarose and acrylamide gels. Methods Enzymol., 152, 61-87 (1987).
  2. ^ Sambrook, Fritsch, and Maniatis (1989) Molecular Cloning: A Laboratory Manual, 2nd ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, volume 3, apendices B.11 and B.23
  3. ^ Stellwagen, E., and Stellwagen, N.C., The free solution mobility of DNA in Tris-acetate-EDTA buffers of different concentrations, with and without added NaCl. Electrophoresis, 23(12), 1935-1941 (2002).
  4. ^ Hayes, V.M. et al., Improvements in gel composition and electrophoretic conditions for broad-range mutation analysis by denaturing gradient gel electrophoresis. Nucleic Acids Res., 27(20), e29 (1999).


[edit] See also

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