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TBE buffer

From Wikipedia, the free encyclopedia

TBE or Tris/Borate/EDTA, is a buffer solution containing a mixture of Tris base, boric acid and EDTA.

In molecular biology, TBE and TAE buffers are often used in procedures involving nucleic acids, the most common being electrophoresis. Tris-acid solutions are effective buffers for slightly basic conditions, which keep DNA deprotonated and soluble in water. EDTA is a chelator of divalent cations, particularly of magnesium (Mg2+). As these ions are necessary co-factors for many enzymes, including contaminant nucleases, the role of the EDTA is to protect the nucleic acids against enzymatic degradation. But since Mg2+ is also a co-factor for many useful DNA-modifying enzymes such as restriction enzymes and DNA polymerases, its concentration in TBE or TAE buffers is generally kept low (typically at around 1 mM).

According to studies by Brody and Kern, sodium boric acid[a] is a superior and cheaper conductive media for most DNA gel electrophoresis applications.[1][2]

Recipe (1 liter of 5X stock solution)

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  • 54 g of Tris base (CAS# 77-86-1, free base)
  • 27.5 g of boric acid (CAS# 10043-35-3)
  • 20 ml of 0.5 M EDTA (CAS# 60-00-4) (pH 8.0)

Adjust pH to 8.3 by HCl.[3]

TBE can be diluted to 1X prior to use in electrophoresis, 0.5x is acceptable as well. Higher concentrations will result in poor results due to excessive heat generation.

See also

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  • LB buffer, lithium borate buffer, a similar buffer containing lithium ions in place of Tris
  • TAE buffer, a similar buffer containing acetic acid in place of boric acid

Notes

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  1. ^ 5 mM disodium borate decahydrate or 10 mM sodium hydroxide, pH adjusted to 8.5 with boric acid.

References

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  1. ^ Brody, J.R.; Kern, S.E. (2004). "History and principles of conductive media for standard DNA electrophoresis" (PDF). Anal Biochem. 333 (1): 1–13. doi:10.1016/j.ab.2004.05.054. PMID 15351274.
  2. ^ 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.
  3. ^ "5X TBE(1.1M Tris; 900mM Borate; 25mM EDTA; pH 8.3)" (PDF). Archived from the original (PDF) on 2017-11-07.