Team:Michigan/Jeremy's Notebook

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(Jeremy's Notebook)
(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.
+
Prepare naphthenic acids stock
-
* Adjust pH of the solution to 10-11 to dissolve naphthenic acids into the solution as sodium salts
+
*Mix 1 g naphthenic acid /L in dilute NaOH.
-
* Making the Growing Broth
+
*Adjust pH of the solution to 10-11 to dissolve naphthenic acids into the solution as sodium salts
-
* Mix 20 ml naphthenic acid stock with 170 ml sterilized water in 500 ml erlenmeyer flask.
+
Making the Growing Broth
-
* Prepare 10 ml  Bushnell-Has medium in vial
+
*Mix 20 ml naphthenic acid stock with 170 ml sterilized water in 500 ml erlenmeyer flask.
-
* Mix arbitrary amount of pseudomonas strain in B-H medium
+
*Prepare 10 ml  Bushnell-Has medium in vial
-
* Transfer medium to erlenmeyer flask
+
*Mix arbitrary amount of pseudomonas strain in B-H medium
-
* Incubation (aerobic conditions)
+
*Transfer medium to erlenmeyer flask
-
* Incubate stock at room temperature (20 °C) on a shaker at 200 rpm.
+
*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


Michigan Header




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.