Terrific Broth Medium

Steven J Burgess, is Sparrow

Published: 2024-07-09 DOI: 10.17504/protocols.io.x54v9pb24g3e/v1

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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.

Steps

Prepare stock solutions

1.

Combine the following components for 1L TB buffer

24g

20g

4mL Glycerol

Make up volume to 900mL with dH2O

Autoclave 121°C 0h 30m 0s

2.

Prepare 1L of 10x TB Salts (0.17 M KH2PO4, 0.72 M K2HPO4) by combining the following

23.1g KH2PO4

125.4g K2HPO4

Make up volume to 1L with dH2O

Autoclave 121°C 0h 30m 0s

Prepare medium

3.

Allow solutions to cool to 60°C then add

100mL of 10x TB salts to 900mL TB buffer.

4.

Store at Room temperature

Practical use

5.

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.

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