TBE buffer

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).

More recently discovered substitutes for TBE and TAE buffers for electrophoresis are available.[1]

Recipe (1 liter of 5X stock solution)

Adjust pH to 8.3 by HCl.[2]

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.

gollark: You... can do that, on APIONET.
gollark: This is neat, it actually manages to make avatars work via webhook.
gollark: Oh, NOW it bridges both ways properly.
gollark: Gibson is bees.
gollark: Okay, so I seem to have accidentally stopped it bridging the other way.

See also
  • 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

References
  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. https://www.eeb.ucla.edu/Faculty/Barber/Web%20Protocols/Protocol7.pdf
This article is issued from Wikipedia. The text is licensed under Creative Commons - Attribution - Sharealike. Additional terms may apply for the media files.