TAE buffer

Source: Wikipedia, the free encyclopedia.

TAE buffer is a

EDTA
.

In molecular biology, it is used in agarose

EDTA, which sequesters divalent cations. TAE has a lower buffer capacity than TBE
and can easily become exhausted, but linear, double stranded DNA runs faster in TAE.

Previously, Brody & Kern simplified electrophoretic buffers by substituting TBE and TAE buffers for a more efficient and inexpensive conductive media in gel systems.[2]

Uses

TAE (Tris-acetate-EDTA) buffer is used as both a running buffer and in agarose gels.[3] Its use in denaturing gradient gel electrophoresis methods for broad-range mutation analysis has also been described.[4] TAE has been used at various concentrations to study the mobility of DNA in solution with and without sodium chloride.[5] However, high concentrations of sodium chloride (and many other salts) in a DNA sample retard its mobility. This may lead to incorrect interpretations of the resulting DNA banding pattern.

Preparation

TAE buffer is commonly prepared as a 50× stock solution for laboratory use. A 50× stock solution can be prepared by dissolving 242 g Tris base in water, adding 57.1 ml glacial acetic acid, and 100 ml of 500 mM EDTA (pH 8.0) solution, and bringing the final volume up to 1 litre. This stock solution can be diluted 49:1 with water to make a 1× working solution. This 1× solution will contain 40 mM Tris, 20 mM acetic acid, and 1 mM EDTA.

No. Name Per 1 mole 50x solution 50× 1× solution 1X
1
Tris base
 121.1 g/L 2 M 242.2 g/L 40 mM 4.844 g/L
2 acetic acid 57.1 ml/L 1 M 57.1 ml/L 20 mM 1.21 ml/L
3
EDTA
disodium salt dihydrate 		372.24 g/L 50 mM 18.612 g/L 1 mM 0.372 g/L

2 M = 2000 mM so 2000 mM /50 = 40 mM for 1×.
1M = 1000 mM so 1000 mM /50 = 20 mM for 1×.
50 mM /50 = 1 mM for 1×.

First of all, these ingredients should be dissolved in 500 ml, then made up to 1000 ml.
Note: EDTA will take more time to dissolve, so while dissolving EDTA use magnetic stirrer (few amounts of EDTA in 3 or 4 times).

A step-by-step recipe of the preparation method for 50× TAE buffer is available on protocols.io.[6]

See also

References

  1. ^ Ogden, R.C., and Adams, D.A., (1987) Electrophoresis in agarose and acrylamide gels. Methods Enzymol., 152:, 61-87.
  2. PMID 15351274 PDF
  3. ISBN 0-87969-309-6
  4. PMID 10497279
  5. PMID 12116139
  6. doi:10.17504/protocols.io.gtvbwn6. {{cite journal}}: Cite journal requires |journal= (help
    )
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