TBE or Tris/Borate/EDTA, is a
buffer solution
A buffer solution (more precisely, pH buffer or hydrogen ion buffer) is an aqueous solution consisting of a mixture of a weak acid and its conjugate base, or vice versa. Its pH changes very little when a small amount of strong acid or base is ...
containing a mixture of
Tris base,
boric acid
Boric acid, more specifically orthoboric acid, is a compound of boron, oxygen, and hydrogen with formula . It may also be called hydrogen borate or boracic acid. It is usually encountered as colorless crystals or a white powder, that dissolve ...
and
EDTA
Ethylenediaminetetraacetic acid (EDTA) is an aminopolycarboxylic acid with the formula H2N(CH2CO2H)2sub>2. This white, water-soluble solid is widely used to bind to iron (Fe2+/Fe3+) and calcium ions (Ca2+), forming water-soluble complexes ev ...
.
In molecular biology, TBE and
TAE buffers are often used in procedures involving
nucleic acids
Nucleic acids are biopolymers, macromolecules, essential to all known forms of life. They are composed of nucleotides, which are the monomers made of three components: a 5-carbon sugar, a phosphate group and a nitrogenous base. The two main cl ...
, the most common being
electrophoresis
Electrophoresis, from Ancient Greek ἤλεκτρον (ḗlektron, "amber") and φόρησις (phórēsis, "the act of bearing"), is the motion of dispersed particles relative to a fluid under the influence of a spatially uniform electric f ...
.
Tris
Tris, or tris(hydroxymethyl)aminomethane, or known during medical use as tromethamine or THAM, is an organic compound with the formula (HOCH2)3CNH2, one of the twenty Good's buffers. It is extensively used in biochemistry and molecular biology a ...
-acid solutions are effective buffers for slightly basic conditions, which keep DNA deprotonated and soluble in water. EDTA is a
chelator
Chelation is a type of bonding of ions and molecules to metal ions. It involves the formation or presence of two or more separate coordinate bonds between a polydentate (multiple bonded) ligand and a single central metal atom. These ligands are ...
of divalent
cations
An ion () is an atom or molecule with a net electrical charge.
The charge of an electron is considered to be negative by convention and this charge is equal and opposite to the charge of a proton, which is considered to be positive by conve ...
, particularly of
magnesium
Magnesium is a chemical element with the symbol Mg and atomic number 12. It is a shiny gray metal having a low density, low melting point and high chemical reactivity. Like the other alkaline earth metals (group 2 of the periodic ...
(Mg
2+). 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 Mg
2+ is also a co-factor for many useful DNA-modifying enzymes such as
restriction enzyme
A restriction enzyme, restriction endonuclease, REase, ENase or'' restrictase '' is an enzyme that cleaves DNA into fragments at or near specific recognition sites within molecules known as restriction sites. Restriction enzymes are one class ...
s and
DNA polymerase
A DNA polymerase is a member of a family of enzymes that catalyze the synthesis of DNA molecules from nucleoside triphosphates, the molecular precursors of DNA. These enzymes are essential for DNA replication and usually work in groups to create ...
s, 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.
Recipe (1 liter of 5X stock solution)
*54 g of
Tris base (CAS# 77-86-1, free base)
*27.5 g of
boric acid
Boric acid, more specifically orthoboric acid, is a compound of boron, oxygen, and hydrogen with formula . It may also be called hydrogen borate or boracic acid. It is usually encountered as colorless crystals or a white powder, that dissolve ...
(CAS# 10043-35-3)
*20 ml of 0.5 M
EDTA
Ethylenediaminetetraacetic acid (EDTA) is an aminopolycarboxylic acid with the formula H2N(CH2CO2H)2sub>2. This white, water-soluble solid is widely used to bind to iron (Fe2+/Fe3+) and calcium ions (Ca2+), forming water-soluble complexes ev ...
(CAS# 60-00-4) (
pH 8.0)
Adjust pH to 8.3 by HCl.
[https://www.eeb.ucla.edu/Faculty/Barber/Web%20Protocols/Protocol7.pdf ]
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
*
LB buffer LB buffer, also known as lithium borate buffer, is a buffer solution used in agarose electrophoresis, typically for the separation of nucleic acids such as DNA and RNA. It is made up of Lithium borate (lithium hydroxide monohydrate and boric acid) ...
, 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
{{DEFAULTSORT:Tbe Buffer
Buffer solutions