Team:Michigan/Jeremy's Notebook

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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.