Apr 06, 2018

Public workspaceRecipe for 50x TAE buffer

DOI
dx.doi.org/10.17504/protocols.io.gtvbwn6
  • Anna Behle1,
  • Alice Pawlowski1
  • 1Institute for Synthetic Microbiology
  • Axmann Lab
  • iGEM Duesseldorf 2018
Anna Behle
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DOI: dx.doi.org/10.17504/protocols.io.gtvbwn6
Protocol Citation: Anna Behle, Alice Pawlowski 2018. Recipe for 50x TAE buffer. protocols.io https://dx.doi.org/10.17504/protocols.io.gtvbwn6
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: Working
We use this protocol and it's working
Created: December 19, 2016
Last Modified: April 06, 2018
Protocol Integer ID: 4693
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
ReagentEDTA
ReagentAcetic acid, glacialMerck MilliporeSigma (Sigma-Aldrich)Catalog #537020
ReagentTris (Tris Base)Gold BiotechnologyCatalog #T-400
Components required
Components required
ComponentMolarity in 50xMolecular weight 
EDTA disodium salt50 mM372.24 g/mol 
Tris2 M 121.14 g/mol 
glacial / acetic acid1 M 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
ad 1000 ml a. dest.
final concentration in gel / running buffer: 40 mM Tris, 20 mM acetic acid, 1 mM EDTA