License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: In development
We are still developing and optimizing this protocol
Created: March 29, 2024
Last Modified: April 30, 2024
Protocol Integer ID: 97568
Abstract
Stock solution for 50x TAE. TAE buffer is a solution made up of Tris base, acetic acid and EDTA (Tris-acetate-EDTA). It is a common buffer for DNA separation using standard agarose gel electrophoresis.
Materials
MATERIALS
EDTA
Acetic acid, glacialSigma AldrichCatalog #537020
Tris (Tris Base)Gold BiotechnologyCatalog #T-400
Components required
Components required
A
B
C
D
E
F
G
Component
mass
VOLUME
Molarity in 50x
Final in 1x
Molecular weight
1
EDTA disodium salt
93.05 g
50 mM
1 mM
372.24 g/mol
na
Dissolve
400 mL
100 mL
pH = 8.0
AUTOCLAVE
2
Tris-base
242 g
2 M
40 mM
121.14 g/mol
na
Dissolve
700 mL
700 mL
3
glacial / acetic acid
57.1 mL
1 M
20 mM
60.05
Preparation of a 0.5 M EDTA stock solution
Preparation of a 0.5 M EDTA stock solution
For 500 ml:
weigh out 93.05 grams of EDTA disodium salt (MW=372.24 g/mol)
Dissolve in 400 milliliter deionized water and adjust the pH with solid sodium hydroxide (NaOH) plates, EDTA will not go completely into solution until the pH is adjusted to about 8.0!
Top up the solution to a final volume of 500 milliliters
autoclave
Preparation of 50 x TAE buffer
Preparation of 50 x TAE buffer
weigh out 242 grams of Tris-base (MW = 121.14 g/mol) and dissolve in approximately 700 milliliters of deionized water
Carefully add 57.1 milliliters of 100 % glacial acid (or acetic acid) and 100 milliliters of 0.5 M EDTA (pH 8.0)
adjust the solution to a final volume of 1 liter
the pH of this buffer is not adjusted and should be about 8.5
store stock solution at room temperature
Preparation of 1 x TAE working solutuion
Preparation of 1 x TAE working solutuion
20 mL 50 x TAE
add 920 mL DI H2O
final concentration in gel / running buffer: 40 mM Tris, 20 mM acetic acid, 1 mM EDTA