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Team:Michigan/Jeremy's Notebook
From 2010.igem.org
(→Jeremy's Notebook) |
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Composition of Medium | Composition of Medium | ||
| - | * 100 mg Naphthenic acids /L | + | *100 mg Naphthenic acids /L |
(Modified Bushnell-Has medium) | (Modified Bushnell-Has medium) | ||
| - | * 1000 mg KH2PO4 /L | + | *1000 mg KH2PO4 /L |
| - | * 1000 mg Na2HPO4 /L | + | *1000 mg Na2HPO4 /L |
| - | * 500 mg NH4NO3 /L | + | *500 mg NH4NO3 /L |
| - | * 500 mg (NH4)2SO4 /L | + | *500 mg (NH4)2SO4 /L |
| - | * 200 mg MgSO4 ∙ 7H2O /L | + | *200 mg MgSO4 ∙ 7H2O /L |
| - | * 20 mg CaCl2 ∙ 2H2O /L | + | *20 mg CaCl2 ∙ 2H2O /L |
| - | * 2 mg FeCl3 /L | + | *2 mg FeCl3 /L |
| - | * 2 mg MnSO4 ∙ 2H2O /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. | + | *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 | + | *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. | + | *Mix 20 ml naphthenic acid stock with 170 ml sterilized water in 500 ml erlenmeyer flask. |
| - | * Prepare 10 ml Bushnell-Has medium in vial | + | *Prepare 10 ml Bushnell-Has medium in vial |
| - | * Mix arbitrary amount of pseudomonas strain in B-H medium | + | *Mix arbitrary amount of pseudomonas strain in B-H medium |
| - | * Transfer medium to erlenmeyer flask | + | *Transfer medium to erlenmeyer flask |
| - | * Incubation (aerobic conditions) | + | *Incubation (aerobic conditions) |
| - | * Incubate stock at room temperature (20 °C) on a shaker at 200 rpm. | + | *Incubate stock at room temperature (20 °C) on a shaker at 200 rpm. |
Revision as of 04:01, 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.
