Clostridium grantii Medium
Clostridium formicoaceticum Agar Composition per liter: Agar ............................................................................................ 15.0g K2HPO4 ....................................................................................... 10.0g Yeast extract.................................................................................. 5.0g Sodium thioglycolate ................................................................. .0.75g Pyridoxine·HCl .......................................................................... 1.0mg Resazurin ................................................................................... 1.0mg Fructose solution......................................................................50.0mL NaHCO3 solution .....................................................................30.0mL Trace elements solution SL-4 ..................................................10.0mL pH 8.0 ± 0.2 at 25°C
Trace Elements Solution SL-4: Composition per liter: EDTA ............................................................................................ 0.5g FeSO4·7H2O................................................................................. 0.2g Trace elements solution SL-6 ................................................100.0mL
Trace Elements Solutions SL-6: Composition per liter: MnCl2·4H2O.................................................................................. 0.5g H3BO3 ........................................................................................... 0.3g CoCl2·6H2O .................................................................................. 0.2g ZnSO4·7H2O ................................................................................ 0.1g Na2MoO4·2H2O ......................................................................... 0.03g NiCl2·6H2O ................................................................................. 0.02g CuCl2·2H2O ................................................................................ 0.01g
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Sodium thioglycolate ................................................................. .0.75g Pyridoxine·HCl .......................................................................... 1.0mg Resazurin ................................................................................... 1.0mg Fructose solution......................................................................50.0mL NaHCO3 solution .....................................................................30.0mL Trace elements solution SL-4 ..................................................10.0mL pH 8.0 ± 0.2 at 25°C
Trace Elements Solution SL-4: Composition per liter: EDTA ............................................................................................ 0.5g FeSO4·7H2O................................................................................. 0.2g Trace elements solution SL-6 ................................................100.0mL
Trace Elements Solution SL-6: Composition per liter: MnCl2·4H2O ................................................................................. 0.5g H3BO3 ........................................................................................... 0.3g CoCl2·6H2O .................................................................................. 0.2g ZnSO4·7H2O ................................................................................ 0.1g Na2MoO4·2H2O ......................................................................... 0.03g NiCl2·6H2O................................................................................. 0.02g CuCl2·2H2O ................................................................................ 0.01g
Preparation of Trace Elements Solution SL-6: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Preparation of Trace Elements Solution SL-4: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly.
to distilled/deionized water and bring volume to 1.0L. Mix thoroughly.
Fructose Solution: Composition per 50.0mL:
Preparation of Trace Elements Solution SL-4: Add components
Fructose......................................................................................... 5.0g
to distilled/deionized water and bring volume to 1.0L. Mix thoroughly.
Preparation of Fructose Solution: Add fructose to distilled/de-
Fructose Solution: Composition per 50.0mL:
ionized water and bring volume to 50.0mL. Mix thoroughly. Sparge under 100% N2 gas for 3 min. Filter sterilize. Store under N2 gas.
Preparation of Trace Elements Solution SL-6: Add components
Preparation of Fructose Solution: Add fructose to distilled/de-
NaHCO3 Solution: Composition per 30.0mL:
ionized water and bring volume to 50.0mL. Mix thoroughly. Sparge under 100% N2 gas for 3 min. Filter sterilize. Store under N2 gas.
NaHCO3 ...................................................................................... 10.0g
Preparation of NaHCO3 Solution: Add NaHCO3 to distilled/de-
NaHCO3 Solution: Composition per 30.0mL:
ionized water and bring volume to 30.0mL. Mix thoroughly. Sparge with 100% CO2. Autoclave for 15 min at 15 psi pressure–121°C.
Fructose......................................................................................... 5.0g
NaHCO3 ...................................................................................... 10.0g
Preparation of NaHCO3 Solution: Add NaHCO3 to distilled/de-
ionized water and bring volume to 30.0mL. Mix thoroughly. Sparge with 100% CO2. Autoclave for 15 min at 15 psi pressure–121°C.
Preparation of Medium: Add components, except fructose solution and NaHCO3 solution, to distilled/deionized water and bring volume to 920.0mL. Mix thoroughly. Gently heat and bring to boiling. Cool to 50°C while sparging with 100% CO2. Autoclave for 15 min at 15 psi pressure–121°C. Aseptically and anaerobically add 50.0mL of sterile fructose solution and 30.0mL of sterile NaHCO3 solution. Mix thoroughly. Adjust pH to 8.0. Aseptically and anaerobically pour into sterile Petri dishes or distribute into sterile screw-capped bottles under 100% CO2. Use: For the cultivation and maintenance of Clostridium formicoaceticum.
Clostridium formicoaceticum Broth Composition per liter: K2HPO4 ....................................................................................... 10.0g Yeast extract.................................................................................. 5.0g © 2010 by Taylor and Francis Group, LLC
Preparation of Medium: Add components, except fructose solution and NaHCO3 solution, to distilled/deionized water and bring volume to 920.0mL. Mix thoroughly. Sparge with 100% CO2. Autoclave for 15 min at 15 psi pressure–121°C. Aseptically and anaerobically add 50.0mL of sterile fructose solution and 30.0mL of sterile NaHCO3 solution. Mix thoroughly. Adjust pH to 8.0. Aseptically and anaerobically distribute into sterile screw-capped bottles under 100% CO2. Use: For the cultivation of Clostridium formicoaceticum.
Clostridium grantii Medium Composition per liter: NaCl.......................................................................................... 19.45g MgCl2............................................................................................ 8.8g Peptone ......................................................................................... 5.0g Na2SO3 ........................................................................................ 3.24g NaHCO3 ........................................................................................ 2.0g CaCl2 ............................................................................................. 1.8g Yeast extract.................................................................................. 1.0g KCl.............................................................................................. 0.55g Ferric citrate.................................................................................. 0.1g