Apr 30, 2024
Recipe for 50x TAE buffer
Forked from Recipe for 50x TAE buffer
This protocol is a draft, published without a DOI.
- Anna Behle1,
- Alice Pawlowski1
- 1Institute for Synthetic Microbiology
- Axmann Lab
- iGEM Duesseldorf 2018
Protocol Citation: Anna Behle, Alice Pawlowski 2024. Recipe for 50x TAE buffer. protocols.io https://dx.doi.org/
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 30, 2024
Last Modified: April 30, 2024
Protocol Integer ID: 97568
Keywords: tae buffer, 50x tae, tae, common buffer, buffer, dna separation, stock solution, agarose gel
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, glacialMerck MilliporeSigma (Sigma-Aldrich)Catalog #537020
Tris (Tris Base)Gold BiotechnologyCatalog #T-400
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
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
- 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
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