Menu
Home
People
Places
Arts
History
Plants & Animals
Science
Life & Culture
Technology
Reference.org
TAE buffer
open-in-new
<h2 id="uses">Uses</h2> <p>TAE (Tris-acetate-EDTA) buffer is used as both a running buffer and in agarose gels.<a class="footnote-ref" id="fnref:4" href="#fn:4"><sup>4</sup></a> Its use in denaturing gradient <a href="/facts/Gel_electrophoresis/9J3GrsFk">gel electrophoresis</a> methods for broad-range mutation analysis has also been described.<a class="footnote-ref" id="fnref:5" href="#fn:5"><sup>5</sup></a> TAE has been used at various concentrations to study the mobility of DNA in solution with and without <a href="/facts/Sodium_chloride/MWcXl3Mb">sodium chloride</a>.<a class="footnote-ref" id="fnref:6" href="#fn:6"><sup>6</sup></a> 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. </p> <h2 id="preparation">Preparation</h2> <p>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. </p> <table><tbody><tr><th>No.</th><th>Name</th><th>Per 1 mole</th><th>50x solution</th><th>50×</th><th>1× solution</th><th>1X</th></tr><tr><td>1</td><td><a href="/facts/Tris_base/m4sonDo5">Tris base</a></td><td>121.1 g/L</td><td>2 M</td><td>242.2 g/L</td><td>40 mM</td><td>4.844 g/L</td></tr><tr><td>2</td><td><a href="/facts/Acetic_acid/uyBKA2Pd">acetic acid</a></td><td>57.1 ml/L</td><td>1 M</td><td>57.1 ml/L</td><td>20 mM</td><td>1.21 ml/L</td></tr><tr><td>3</td><td><a href="/facts/EDTA/XvHgKQyD">EDTA</a> disodium salt dihydrate</td><td>372.24 g/L</td><td>50 mM</td><td>18.612 g/L</td><td>1 mM</td><td>0.372 g/L</td></tr></tbody></table> <p>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×. </p><p>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). </p><p>A step-by-step recipe of the preparation method for 50× TAE buffer is available on protocols.io.<a class="footnote-ref" id="fnref:7" href="#fn:7"><sup>7</sup></a> </p> <h2 id="see-also">See also</h2> <ul><li><a href="/facts/TBE_buffer/RmYhCdGd">TBE buffer</a></li> <li><a href="/facts/LB_buffer/cJK9T6TW">LB buffer</a></li></ul> <h2 id="references">References</h2> <ol> <li id="fn:1"><p>Ogden, R.C., and Adams, D.A., (1987) Electrophoresis in agarose and acrylamide gels. Methods Enzymol., 152:, 61-87. <a href="#fnref:1" class="footnote-back-ref">↩</a></p></li> <li id="fn:2"><p>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 <a href="/wiki/Doi_(identifier)" target="_blank">/wiki/Doi_(identifier)</a> <a href="#fnref:2" class="footnote-back-ref">↩</a></p></li> <li id="fn:3"><p>Brody, Jonathan R.; Kern, Scott E. (February 2004). "Sodium boric acid: a Tris-free, cooler conductive medium for DNA electrophoresis". BioTechniques. 36 (2): 214–216. doi:10.2144/04362BM02. <a href="/wiki/Doi_(identifier)" target="_blank">/wiki/Doi_(identifier)</a> <a href="#fnref:3" class="footnote-back-ref">↩</a></p></li> <li id="fn:4"><p>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 <a href="/wiki/ISBN_(identifier)" target="_blank">/wiki/ISBN_(identifier)</a> <a href="#fnref:4" class="footnote-back-ref">↩</a></p></li> <li id="fn:5"><p>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 <a href="/wiki/PMID_(identifier)" target="_blank">/wiki/PMID_(identifier)</a> <a href="#fnref:5" class="footnote-back-ref">↩</a></p></li> <li id="fn:6"><p>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 <a href="/wiki/PMID_(identifier)" target="_blank">/wiki/PMID_(identifier)</a> <a href="#fnref:6" class="footnote-back-ref">↩</a></p></li> <li id="fn:7"><p>Behle, Anna; Pawlowski, Alice (6 April 2018). "Recipe for 50x TAE buffer". doi:10.17504/protocols.io.gtvbwn6. {{cite journal}}: Cite journal requires |journal= (help) <a href="https://www.protocols.io/view/recipe-for-50x-tae-buffer-gtvbwn6" target="_blank">https://www.protocols.io/view/recipe-for-50x-tae-buffer-gtvbwn6</a> <a href="#fnref:7" class="footnote-back-ref">↩</a></p></li> </ol>