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From 2010.igem.org

Jeremy's Notebook

Growing Broth for Pseudomonas

Composition of Medium *100 mg Naphthenic acids /L

(Modified Bushnell-Has medium) *1000 mg KH2PO4 /L *1000 mg Na2HPO4 /L *500 mg NH4NO3 /L *500 mg (NH4)2SO4 /L *200 mg MgSO4 ∙ 7H2O /L *20 mg CaCl2 ∙ 2H2O /L *2 mg FeCl3 /L *2 mg MnSO4 ∙ 2H2O /L pH of Bushnell-Has Medium: 7.0 Protocol Prepare naphthenic acids stock *Mix 1 g naphthenic acid /L in dilute NaOH. *Adjust pH of the solution to 10-11 to dissolve naphthenic acids into the solution as sodium salts Making the Growing Broth *Mix 20 ml naphthenic acid stock with 170 ml sterilized water in 500 ml erlenmeyer flask. *Prepare 10 ml Bushnell-Has medium in vial *Mix arbitrary amount of pseudomonas strain in B-H medium *Transfer medium to erlenmeyer flask *Incubation (aerobic conditions) *Incubate stock at room temperature (20 °C) on a shaker at 200 rpm.