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Team:Michigan/Jeremy's Notebook
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Composition of Medium | Composition of Medium | ||
| - | + | * 100 mg Naphthenic acids /L | |
(Modified Bushnell-Has medium) | (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 | pH of Bushnell-Has Medium: 7.0 | ||
Protocol | 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. | |
Revision as of 03:59, 18 July 2010
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.
