Jan 25, 2016
TE Buffer
- Ms Alex Aitken
- VERVE Net
- Sullivan Lab
Protocol Citation: Ms Alex Aitken 2016. TE Buffer. protocols.io https://dx.doi.org/10.17504/protocols.io.c5uy6v
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
Created: June 22, 2015
Last Modified: March 14, 2018
Protocol Integer ID: 916
Abstract
TE is a commonly used buffer solution in molecular biology, especially in procedures involving DNA or RNA. "TE" is derived from its components: Tris, a common pH buffer, and EDTA, a molecule that chelates cations like Mg2+. The purpose of TE buffer is to solubilize DNA or RNA, while protecting it from degradation: 10 mM Tris, bring to pH 8.0 with HCl, 1 mM EDTA
Guidelines
| Component | For 500ml | |
| 1M Tris pH 8* | 5ml | |
| 0.5M EDTA pH 8 | 1ml | |
| dH2O | 496ml |
*the pH is usually adjusted to RNA 8.0 for DNA and 7.5 forThe respective DNA and RNA nucleases are supposed to be less active at these pH values, but pH 8.0 can safely be used for storage of both DNA and RNA. EDTA further inactivates nucleases, by binding to metal cations required by these enzymes. Genomic and plasmid DNA can be stored in TE Buffer at 4ºC (39.2ºF) for short-term use, or -20ºC (-4ºF) to -80ºC (-112ºF) for long-term storage.
Safety warnings
Repeated freeze-thaw cycles should be avoided
Prepare 5ml of 1M Tris
61 g
Protocol
NAME
1M TrisCREATED BY
Verve Team
60.57 g of Tris (hydroxymethyl) aminomethane in 0.5L Milli-Qwater
Bring pH to 8.0 using HCl
Making 0.5M EDTA
Making 0.5M EDTA
Prepare 1ml 0.5M EDTA
19 g
Protocol
NAME
0.5M EDTACREATED BY
Verve Team
Mix 18.6 g EDTA in 100ml Milli-Q water
19 g
Bring pH to 8.0 using NaOH
Note
EDTA will not be soluble until pH reaches 8.0–this will take time (hours)
Combine 5ml of 1M Tris and 1ml of 0.5M EDTA in 496ml dH2O
Autoclave to sterilize.
Note
It is best to remove a working aliquot and not repeatedly access the stock