Methanomicrobium Medium

[所属分类:培养基配方] [发布时间:2021-5-26] [发布人:网站管理员2] [阅读次数:] [返回]

Methanomicrobium Medium

山东拓普生物工程有限公司 培养基配方 http://www.topbiol.com

Composition per liter:

NaHCO 3 ........................................................................................2.0g
Yeast extract..................................................................................1.0g
Pancreatic digest of casein............................................................1.0g
NaCl..............................................................................................0.6g
L- Cysteine·HCl·H 2 O......................................................................0.5g
Na 2 S·9H 2 O....................................................................................0.5g
K 2 HPO 4 .........................................................................................0.3g
KH 2 PO 4 .........................................................................................0.3g
(NH 4 ) 2 SO 4 .....................................................................................0.3g
MgSO 4 ·7H 2 O..............................................................................0.13g
CaCl 2 ·2H 2 O................................................................................8.0mg
FeSO 4 ·7H 2 O...............................................................................2.0mg
Rumen fluid, clarified............................................................300.0mL
Fatty acid mixture....................................................................20.0mL
Wolfe’s mineral solution..........................................................10.0mL
Wolfe’s vitamin solution..........................................................10.0mL
Resazurin (0.1% solution) .........................................................1.0mL
pH 6.5 ± 0.2 at 25°C
Fatty Acid Mixture:
Composition per liter:
Valeric acid................................................................................0.7mL
Isovaleric acid............................................................................0.7mL
α-Methylbutyric acid.................................................................0.5mL
Isobutyric acid ...........................................................................0.5mL
Preparation of Fatty Acid Mixture: Add components to dis-
tilled/deionized water and bring volume to 1.0L. Mix thoroughly.
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
FeSO 4 ·7H 2 O..................................................................................0.1g
CoCl 2 ·6H 2 O..................................................................................0.1g
CaCl 2 .............................................................................................0.1g
ZnSO 4 ·7H 2 O.................................................................................0.1g
CuSO 4 ·5H 2 O...............................................................................0.01g
AlK(SO 4 ) 2 ·12H 2 O.......................................................................0.01g
H 3 BO 3 .........................................................................................0.01g
Na 2 MoO 4 ·2H 2 O..........................................................................0.01g
Preparation of Wolfe’s Mineral 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.
Wolfe’s Vitamin Solution:
Composition per liter:
Pyridoxine·HCl........................................................................10.0mg
Thiamine·HCl............................................................................5.0mg
Riboflavin..................................................................................5.0mg
Nicotinic acid.............................................................................5.0mg
Calcium pantothenate ................................................................5.0mg
p-Aminobenzoic acid.................................................................5.0mg
Thioctic acid..............................................................................5.0mg
Biotin.........................................................................................2.0mg
Folic acid ...................................................................................2.0mg
Cyanocobalamin.....................................................................100.0μg
Preparation of Wolfe’s Vitamin Solution: Add components to
distilled/deionized water and bring volume to 1.0L. Mix thoroughly.
Preparation of Medium:  Prepare and dispense medium under 80%
N 2 + 20% CO 2 . Add components to distilled/deionized water and bring
volume to 1.0L. Mix thoroughly. Adjust pH to 6.5. Distribute into tubes
or flasks under 80% N 2 + 20% CO 2 . Cap with rubber stoppers. Auto-
clave for 15 min at 15 psi pressure–121°C.
Use:  For the cultivation and maintenance of Methanomicrobium
mobile.