Jul 09, 2024
Terrific Broth Medium
- is Sparrow1,
- Steven Burgess1
- 1UIUC
Protocol Citation: is Sparrow, Steven Burgess 2024. Terrific Broth Medium. protocols.io https://dx.doi.org/10.17504/protocols.io.x54v9pb24g3e/v1
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: January 03, 2024
Last Modified: July 09, 2024
Protocol Integer ID: 92922
Keywords: growing plasmid, cosmid clone, terrific broth medium tartoff, improved media, laboratory
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Abstract
Tartoff, K.D. and Hobbs, C.A. (1987) Improved Media for Growing Plasmid and Cosmid Clones. Bethesda Research Laboratories Focus, 9, 12.
Guidelines
Prepare the stock solutions and autoclave separately before combining when cooled to create TB medium. The reason for this is that Glycerol is believed to react with amines in the Tryptone/Yeast extract via the Malliard reaction, leading to the formation of compounds which
Protocol materials
Bacto™ Tryptone Life TechnologiesVWR International (Avantor)Catalog #211705
BD Bacto™ Yeast Extract Becton Dickinson (BD)Catalog #212750
Prepare stock solutions
1h
Combine the following components for 1L TB buffer
24 g BD Bacto™ Yeast Extract Becton Dickinson (BD)Catalog #212750
20 g Bacto™ Tryptone Life TechnologiesVWR International (Avantor)Catalog #211705
4 mL Glycerol
Make up volume to 900 mL with dH2O
Autoclave 121 °C 00:30:00
30m
Prepare 1L of 10x TB Salts (0.17 M KH2PO4, 0.72 M K2HPO4) by combining the following
23.1 g KH2PO4
125.4 g K2HPO4
Make up volume to 1 L with dH2O
Autoclave 121 °C 00:30:00
30m
Prepare medium
1h
Allow solutions to cool to 60 °C or less then add
100 mL of 10x TB salts to 900 mL TB buffer.
Store at Room temperature
Practical use
We have found that using TB for plasmid preps consistently yields higher than using LB regardless of prep kit used, with half the volume of culture required.
It is not recommended to exceed ~2 mL of saturated TB culture per prep column due to overloading the column and not yielding clean preps.