Cell Culture Microbiology

scientificprotocols authored over 3 years ago

Authors: Mark Dopson & Stephan Christel

Abstract

Medium for growth of extreme acidophilic microorganisms. This medium does not contain trace elements. When not working on mineral, addition of trace element (TE) solution is necessary, see separate protocol. http://www.nature.com/protocolexchange/protocols/3811

Reagents

Medium:

  1. Ammonium sulfate, (NH4)2SO4
  2. Sodium sulfate decahydrate, Na2SO4 x10H2O
  3. Potassium chloride, KCl
  4. Potassium hydrogen phosphate, K2HPO4
  5. Magnesium sulfate heptahydrate, MgSO4 x7H2O
  6. Calcuim nitrate, Ca(NO3)2

General:

  1. Sulfuric acid
  2. Sodium hydroxide

Equipment

  1. Stirring rods
  2. Magnetic stirrer
  3. Measuring colve
  4. pH meter

Procedure

  • For 1x Medium:

    • 3.0 g/l (NH4)2SO4
    • 3.2 g/l Na2SO4 x10H2O
    • 0.1 g/l KCl
    • 0.05 g/l K2HPO4
    • 0.5 g/l MgSO4 x7H2O
    • 0.01 g/l Ca(NO3)2
  • For 10x Stock:

    • 30 g/l (NH4)2SO4
    • 32 g/l Na2SO4 x10H2O
    • 1 g/l KCl
    • 0.5 g/l K2HPO4
    • 5 g/l MgSO4 x7H2O
    • 0.1 g/l Ca(NO3)2
  • Add the constituents into a final volume of 1 liter of water, one at a time, dissolving before adding the next.

  • Adjust the pH to 2.0 with sulfuric acid (and/or sodium hydroxide)

  • Autoclave

Timing

15 minutes + autoclaving time

Author information

Mark Dopson & Stephan Christel, Systems Biology of Extreme Microorganisms, Dopson, Linnaeus University

Correspondence to: Stephan Christel ([email protected])

Source: Protocol Exchange (2015) doi:10.1038/protex.2015.031. Originally published online 24 March 2015.

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