In molecular biology it is 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 8.3, and 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.
Recently, Brody & Kern simplified electrophoretic buffers by substituting TBE and TAE buffers for a more efficient and inexpensive conductive media in gel systems.
TAE (Tris-acetate-EDTA) buffer is used as both a running buffer and in agarose gel. Its use in denaturing gradient gel electrophoresis methods for broad-range mutation analysis has also been described. TAE has been used at various concentrations to study the mobility of DNA in solution with and without sodium chloride. 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.
TAE buffer is commonly prepared as a 50X stock solution for laboratory use. A 50X stock solution can be prepared by dissolving 242g Tris base in water, adding 57.1mL glacial acetic acid, and 100mL of 500mM 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 1X working solution. This 1X solution will contain 40mM Tris, 20mM acetic acid, and 1mM EDTA.
|S.No||Name||Per 1 Mole||Main Composition||50X||1X Composition||1X|
|1||Tris Base||121.1 g/l||2M/l||242.2 g/l||40mM||4.844 g/l|
|2||Acetic Acid||60.5 ml/l||1M/l||60.5 ml/l||20mM||1.21 ml/l|
|3||EDTA||372.24 g/l||50 mM/l||18.612 g/l||1mM||0.372 g/l|
2M = 2000 mM so 2000/50 = 40mM for 1x.
1M = 1000mM so 1000/50 = 20mM for 1x.
50mM/l in 50x 50/50 = 1mM for 1x.
first of all, these ingredients should dissolve in 500ml then makeup to 1000 ml
Note: EDTA will take more time to dissolve, so while dissolving EDTA use Magnetic Stirrer (few amounts of EDTA in 3 - 4 times)
- Ogden, R.C., and Adams, D.A., (1987) Electrophoresis in agarose and acrylamide gels. Methods Enzymol., 152:, 61-87.
- Brody, J.R., Kern, S.E. (2004) History and principles of conductive media for standard DNA electrophoresis. Anal Biochem. 333(1):1-13. doi:10.1016/j.ab.2004.05.054 PMID 15351274 PDF
- 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 ISBN 0-87969-309-6
- Hayes, V.M. et al., (1999) Improvements in gel composition and electrophoretic conditions for broad-range mutation analysis by denaturing gradient gel electrophoresis. Nucleic Acids Res., 27(20): e29. PMID 10497279
- Stellwagen, E., and Stellwagen, N.C. (2002) The free solution mobility of DNA in Tris-acetate-EDTA buffers of different concentrations, with and without added NaCl. Electrophoresis, 23(12): 1935-1941. PMID 12116139