Geobacter Medium (DSMZ Medium 826)

[所属分类:培养基配方] [发布时间:2021-3-10] [发布人: ] [阅读次数:] [返回]

Geobacter Medium

(DSMZ Medium 826)

培养基配方

Composition per 1050.0mL:
NaHCO 3 ........................................................................................2.5g
NH 4 Cl ...........................................................................................1.5g
Na-acetate ...................................................................................0.82g
Na 2 HPO 4 .......................................................................................0.6g
KCl................................................................................................0.1g
Resazurin ...................................................................................0.5mg
Na-fumarate solution ...............................................................50.0mL
Trace elements solution ...........................................................10.0mL
Vitamin solution.......................................................................10.0mL
Selenite-tungstate solution.........................................................1.0mL
pH 6.8 ± 0.2 at 25°C
Na-Fumarate Solution:
Composition per 100.0mL:
Na-fumarate................................................................................16.0g
Preparation of Na-Fumarate Solution:  Add Na-fumarate to distilled/deionized water and bring volume to 100.0mL. Mix thoroughly. Sparge with 100% N 2 . Autoclave for 15 min at 15 psi pressure–121°C.
Selenite-Tungstate Solution
Composition per liter:
NaOH............................................................................................0.5g
Na 2 WO 4 ·2H 2 O...........................................................................4.0mg
Na 2 SeO 3 ·5H 2 O...........................................................................3.0mg
Preparation of Selenite-Tungstate Solution: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Sparge with 100% N 2 . Filter sterilize.
Trace Elements Solution:
Composition per liter:
MgSO 4 ·7H 2 O................................................................................3.0g
Nitrilotriacetic acid.......................................................................1.5g
NaCl..............................................................................................1.0g
MnSO 4 ·2H 2 O................................................................................0.5g
CoSO 4 ·7H 2 O...............................................................................0.18g
ZnSO 4 ·7H 2 O...............................................................................0.18g
CaCl 2 ·2H 2 O...................................................................................0.1g
FeSO 4 ·7H 2 O..................................................................................0.1g
NiCl 2 ·6H 2 O...............................................................................0.025g
KAl(SO 4 ) 2 ·12H 2 O.......................................................................0.02g
H 3 BO 3 .........................................................................................0.01g
Na 2 MoO 4 ·4H 2 O ..........................................................................0.01g
CuSO 4 ·5H 2 O...............................................................................0.01g
Na 2 SeO 3 ·5H 2 O...........................................................................0.3mg
Preparation of Trace Elements Solution:  Add nitrilotriacetic acid to 500.0mL of distilled/deionized water. Dissolve by adjusting pH to 6.5 with KOH. Add remaining components. Add distilled/deionized water to 1.0L. Mix thoroughly.

Vitamin Solution:
Composition per liter:
Pyridoxine-HCl........................................................................10.0mg
Thiamine-HCl·2H 2 O..................................................................5.0mg
Riboflavin..................................................................................5.0mg
Nicotinic acid.............................................................................5.0mg
D -Ca-pantothenate .....................................................................5.0mg
p-Aminobenzoic acid.................................................................5.0mg
Lipoic acid.................................................................................5.0mg
Biotin.........................................................................................2.0mg
Folic acid ...................................................................................2.0mg
Vitamin B 12 ................................................................................0.1mg
Preparation of Vitamin Solution:  Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Sparge with 80% H 2 + 20% CO 2 . Filter sterilize.
Preparation of Medium: Prepare and dispense medium under 80% N 2 + 20% CO 2  gas mixture. Add components, except NaHCO 3 and fumarate solution, to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently heat and bring to boiling. Boil for 5 min. Cool to room temperature while sparging with 80% N 2 + 20% CO 2 . Add the NaHCO 3 . Equilibrate with the 80% N 2 + 20% CO 2  gas mixture to reach a pH of 6.8. Distribute into anaerobe tubes under 80% N 2 + 20% CO 2 . Autoclave for 15 min at 15 psi pressure–121°C. Cool to room temperature. Aseptically and anaerobically add 0.5mL Na-fumarate solution per 10.0mL medium. The pH should be 6.8.
Use:  For the cultivation of Geobacter sulfurreducens, Geobacter bremensis (Geobacter sp.), Geobacter chapellei, Geobacter hydrogenophilus, and Geothrix fermentans.