May 14, 2018
LB (Luria-Bertani) liquid medium
- 1University of Edinburgh
- Vale Lab
- Molecular Biology
Protocol Citation: Katy M Monteith 2018. LB (Luria-Bertani) liquid medium. protocols.io https://dx.doi.org/10.17504/protocols.io.p4vdqw6
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: May 14, 2018
Last Modified: May 14, 2018
Protocol Integer ID: 12149
Keywords: LB broth, Luria-Bertani, liquid media
Abstract
This protocol decribes how to make 1 L of Luria-Bertani liquid medium (also known as LB broth) for the culturing of bacteria.
Guidelines
LB broth can be stored at room temperature once autoclaved and kept for up to 6 months.
If your broth becomes cloudy it has become contaminated and is no longer viable. Contaminated LB broth must be disinfected (i.e. by adding Virkon at 2% final concentration) and then autoclaved before disposal.
Materials
MATERIALS
LB broth, MillerThermo Fisher ScientificCatalog #BP1426-500
Virkon powderVWR International (Avantor)Catalog #148-0201
Safety warnings
Read appropriate SDS for LB broth, Miller and Virkon before use.
Bottle can be extremely hot immediately following autoclave. Use thermal gloves for handling.
Add 25 grams of LB broth, Miller per litre of distilled H₂O.
Mix well by inverting bottle several times until powder dissolved.
Sterilize by autoclaving for 20 min at 15 psi (1.05 kg/cm2) on liquid cycle.
Note
Once sterilized only open and use LB broth within a laminar flow hood to prevent contamination.
Allow LB broth to cool to room temperature before use.