Mar 11, 2026

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. protocols.io https://dx.doi.org/
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 11, 2026
Last Modified: March 11, 2026
Protocol  Integer ID: 313119
Keywords: recipe lysogeny broth, lysogeny broth, lb broth, used medium in microbiology, microbiology, catabolizable amino acid, lb, media preparation, 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
Collect Glassware
Collect a 1 L beaker.

Ensure the beaker is clean by rinsing with DI H2O
Combine Media
To prepare 1 liter, dissolve 10 g tryptone, 5 g yeast extract, and 10 g NaCl in 950 mL of deionized water and fill to 1 L
Optional: Aliquot LB
Aliquot LB into (4) 250 mL bottles
Autoclave
Autoclave at 121°C for 20-30 minutes
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.