ProtocolsRecipesLysogenyBrothAgar < Lab

---+++ 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 directly combined with the other components and 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.

Barrick Lab > ProtocolsMediaRecipes > ProtocolsRecipesLysogenyBrothAgar

Topic revision: r11 - 2025-02-19 - 19:00:49 - Main.TylerDeJong

Lab.ProtocolsRecipesLysogenyBrothAgar moved from Lab.ProtocolsRecipesLysogenyBroth on 2015-06-23 - 14:17 by Main.JeffreyBarrick -