Mar 12, 2026

Public workspaceLysogeny broth (LB) Media Preparation and Recipe_v1

 Forked from Lysogeny broth (LB) Media Preparation and Recipe
This protocol is a draft, published without a DOI.
  • Tamra Lahom1
  • 1Carleton College
Tamra Mbeuh Lahom Lot
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Protocol Citation: Tamra Lahom 2026. Lysogeny broth (LB) Media Preparation and Recipe_v1. protocols.io https://protocols.io/view/lysogeny-broth-lb-media-preparation-and-recipe-v1-jv2acn8af
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: March 12, 2026
Last Modified: March 12, 2026
Protocol Integer ID: 313122
Keywords: lysogeny broth, recipe-v1 lysogeny broth, lb broth, used medium in microbiology, microbiology, catabolizable amino acid, media preparation, lb, bertani broth, carbon source
Abstract
Lysogeny broth (LB) media is one the most used medium in microbiology. LB broth supports Escherichia coli growth to an optical density at 600 nm (OD600) of 7. The carbon sources for E. coli in Luria-Bertani broth are catabolizable amino acids, not sugars.
Materials
  1. 1 L Beaker
  2. Tryptone,
  3. Yeast extract
  4.  NaCl
  5. Deionized water
  6. Autoclave
Troubleshooting
Collect Glassware
Collect a Amount1 L beaker.

Ensure the beaker is clean by rinsing with DI H2O
Combine Media
To prepare 1 L, dissolve Amount10 g tryptone, Amount5 g yeast extract, and Amount10 g NaCl in Amount0.950 L of deionized water and fill to Amount1 L
Note
Have a little water in the bottle before you add the dry powder.

Optional: Aliquot LB
Aliquot LB into (4) 250 mL bottles
Autoclave
Place autoclave tape on the lid of the bottle and autoclave at 121°C for 20-30 minutes
Note
Make sure the lid is loosened to avoid pressure build up.

Storage
Store at room temperature or in a fridge until use.
Protocol references
Sezonov G, Joseleau-Petit D, D'Ari R. Escherichia coli physiology in Luria-Bertani broth. J Bacteriol. 2007 Dec;189(23):8746-9. doi: 10.1128/JB.01368-07. Epub 2007 Sep 28. PMID: 17905994; PMCID: PMC2168924.