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Team:Michigan/Jeremy's Notebook
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Growing Broth for Pseudomonas | Growing Broth for Pseudomonas | ||
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 | ||
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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. | ||
Latest revision as of 04:10, 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.
