Geobacter Medium

[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

培养基配方

Composition per liter:
NaHCO 3 ........................................................................................2.5g
NH 4 Cl ...........................................................................................1.5g
Sodium acetate............................................................................0.82g
NaH 2 PO 4 .......................................................................................0.6g
KCl................................................................................................0.1g
Sodium fumarate solution........................................................50.0mL
Modified Wolfe’s mineral solution..........................................10.0mL
Wolfe’s vitamin solution..........................................................10.0mL
pH 6.8 ± 0.2 at 25°C
Sodium Fumarate Solution:
Composition per 50.0mL:
Sodium fumarate...........................................................................8.0g
Preparation of Sodium Fumarate Solution:  Add sodium fumarate to distilled/deionized water and bring volume to 50.0mL. Mix thoroughly. Filter sterilize. Sparge with 100% N 2 .
Wolfe’s Vitamin Solution:
Composition per liter:
Pyridoxine·HCl........................................................................10.0mg
p-Aminobenzoic acid.................................................................5.0mg
Lipoic acid .................................................................................5.0mg
Nicotinic acid.............................................................................5.0mg
Riboflavin ..................................................................................5.0mg
Thiamine·HCl ............................................................................5.0mg
Calcium  DL -pantothenate...........................................................5.0mg
Biotin .........................................................................................2.0mg
Folic acid....................................................................................2.0mg
Vitamin B 12 ................................................................................0.1mg
Preparation of Wolfe’s Vitamin Solution:  Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly.

Modified Wolfe’s Mineral Solution:
Composition per liter:
MgSO 4 ·7H 2 O................................................................................3.0g
Nitrilotriacetic acid.......................................................................1.5g
NaCl..............................................................................................1.0g
MnSO 4 ·H 2 O..................................................................................0.5g
CaCl 2 .............................................................................................0.1g
CoCl 2 ·6H 2 O..................................................................................0.1g
FeSO 4 ·7H 2 O..................................................................................0.1g
ZnSO 4 ·7H 2 O.................................................................................0.1g
AlK(SO 4 ) 2 ·12H 2 O.......................................................................0.01g
CuSO 4 ·5H 2 O...............................................................................0.01g
H 3 BO 3 .........................................................................................0.01g
Na 2 MoO 4 ·2H 2 O..........................................................................0.01g
Na 2 SeO 3 ......................................................................................0.01g
NaWO 4 ·2H 2 O .............................................................................0.01g
NiC1 2 ·6H 2 O................................................................................0.01g
Preparation of Modified Wolfe’s Mineral Solution: Add nitrilotriacetic acid to 500.0mL of distilled/deionized water. Adjust pH to 6.5 with KOH. Add remaining components one at a time. Add distilled/deionized water to 1.0L. Adjust pH to 6.8.
Preparation of Medium: Prepare and dispense medium under 80% N 2 + 20% CO 2 . Add components, except sodium fumarate solution, to distilled/deionized water and bring volume to 950.0mL. Mix thoroughly. Gently heat and bring to boiling. Continue boiling for 3 min. Cool to room temperature while sparging with 80% N 2 + 20% CO 2 . Anaerobically distribute 9.5mL volumes into anaerobic tubes. Autoclave for 15 min at 15 psi pressure–121°C. Aseptically and anaerobically add 0.5mL of sterile sodium fumarate solution to each tube. Mix thoroughly. Adjust pH to 6.8.
Use: For the cultivation of Geobacter sulfurreducens.