TAE buffer: Difference between revisions
imported>Nosferattus sodium boric acid |
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In molecular biology, it is used in agarose [[electrophoresis]] typically for the separation of [[nucleic acids]] such as [[DNA]] and [[RNA]].<ref>Ogden, R.C., and Adams, D.A., (1987) Electrophoresis in agarose and acrylamide gels. ''Methods Enzymol''., '''152''':, 61-87.</ref> 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 buffer|TBE]] and can easily become exhausted, but linear, double stranded DNA runs faster in TAE. | In molecular biology, it is used in agarose [[electrophoresis]] typically for the separation of [[nucleic acids]] such as [[DNA]] and [[RNA]].<ref>Ogden, R.C., and Adams, D.A., (1987) Electrophoresis in agarose and acrylamide gels. ''Methods Enzymol''., '''152''':, 61-87.</ref> 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 buffer|TBE]] and can easily become exhausted, but linear, double stranded DNA runs faster in TAE. | ||
According to studies by Brody and Kern, sodium boric acid{{efn|5 mM disodium borate decahydrate or 10 mM sodium hydroxide, pH adjusted to 8.5 with boric acid.}} is a superior and cheaper conductive media for most DNA [[gel electrophoresis]] applications.<ref>{{cite journal | last1 = Brody | first1 = J.R. | last2 = Kern | first2 = S.E. | year = 2004 | title = History and principles of conductive media for standard DNA electrophoresis | url = http://www.cc.ahs.chula.ac.th/Molmed/DNA%20electro%27s%20conductive%20media.pdf | journal = Anal Biochem | volume = 333 | issue = 1| pages = 1–13 | doi = 10.1016/j.ab.2004.05.054 | pmid=15351274}}</ref><ref name="BrodyKern2004">{{cite journal |last1=Brody |first1=Jonathan R. |last2=Kern |first2=Scott E. |title=Sodium boric acid: a Tris-free, cooler conductive medium for DNA electrophoresis |journal=BioTechniques |date=February 2004 |volume=36 |issue=2 |pages=214–216 |doi=10.2144/04362BM02}}</ref> | According to studies by Brody and Kern, sodium boric acid{{efn|5 mM disodium borate decahydrate or 10 mM sodium hydroxide, pH adjusted to 8.5 with boric acid.}} is a superior and cheaper conductive media for most DNA [[gel electrophoresis]] applications.<ref>{{cite journal | last1 = Brody | first1 = J.R. | last2 = Kern | first2 = S.E. | year = 2004 | title = History and principles of conductive media for standard DNA electrophoresis | url = http://www.cc.ahs.chula.ac.th/Molmed/DNA%20electro%27s%20conductive%20media.pdf | journal = Anal Biochem | volume = 333 | issue = 1| pages = 1–13 | doi = 10.1016/j.ab.2004.05.054 | pmid=15351274}}</ref><ref name="BrodyKern2004">{{cite journal |last1=Brody |first1=Jonathan R. |last2=Kern |first2=Scott E. |title=Sodium boric acid: a Tris-free, cooler conductive medium for DNA electrophoresis |journal=BioTechniques |date=February 2004 |volume=36 |issue=2 |pages=214–216 |doi=10.2144/04362BM02|doi-access=free }}</ref> | ||
==Uses== | ==Uses== | ||
Latest revision as of 05:03, 24 June 2025
Template:Short description TAE buffer is a buffer solution containing a mixture of Tris base, acetic acid and EDTA.
In molecular biology, it is 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 8.3, 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.
According to studies by Brody and Kern, sodium boric acidTemplate:Efn is a superior and cheaper conductive media for most DNA gel electrophoresis applications.[2][3]
Uses
TAE (Tris-acetate-EDTA) buffer is used as both a running buffer and in agarose gels.[4] Its use in denaturing gradient gel electrophoresis methods for broad-range mutation analysis has also been described.[5] TAE has been used at various concentrations to study the mobility of DNA in solution with and without sodium chloride.[6] 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.[7]
See also
Notes
References
- ↑ Ogden, R.C., and Adams, D.A., (1987) Electrophoresis in agarose and acrylamide gels. Methods Enzymol., 152:, 61-87.
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- ↑ 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 Template:ISBN
- ↑ 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. Template:Catalog lookup link
- ↑ 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. Template:Catalog lookup link
- ↑ Script error: No such module "Citation/CS1".