---+++ LB: Lysogeny Broth / Luria-Bertani (Miller) Agar Add dH<sub>2</sub>O to full volume in an appropriately-sized flask. Combine the following, making sure each component fully dissolves before adding the next: | *1L* | *6L* | *Component* | | 10 g | 60 g | Tryptone | | 5 g | 30 g | Yeast Extract | | 10 g | 60 g | <nop>NaCl | | 1 ml | 6 ml | 5% Antifoam* | | 16 g | 48 g | Agar* | *ANTIFOAM:* Open the bottle under sterile conditions (near a flame) only, and continue agitating after the addition of the 5% antifoam. *AGAR:* Measure 8 grams of agar per 500 ml bottle and *add to each individual bottle before aliquoting* the LB. Autoclave to homogenize. Each liter of LB Agar makes 2 bottles of 500 ml. *Notes and variations:* * [[http://en.wikipedia.org/wiki/Lysogeny_broth][Common synonyms]] are Luria-Broth and Luria-Bertani medium. * This is the "Miller" formulation of LB with 10 g <nop>NaCl per liter (1% w/v). *Another common formulation has only 5 g <nop>NaCl per liter (0.5% w/v)!* They are NOT interchangeable. * If you are performing _sacB_ counter-selection, you do not want to include NaCl in your agar. Use the above recipe, but swap out 5-10% w/v (50-100g per liter) sucrose in place of any NaCl. Sucrose can be autoclaved (no need to filter sterilize it separately). * To make "soft" or "top" agar for phage studies, use only 7-8 g agar per liter.
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Topic revision: r10 - 2025-02-18 - TylerDeJong