Methanosarcina acetivorans 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]

Methanosarcina acetivorans 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:

NaCl............................................................................................23.4g
Agar ............................................................................................10.0g
MgSO 4 ..........................................................................................6.3g
Na 2 CO 3 .........................................................................................5.0g
Trimethylamine·HCl.....................................................................3.0g
Yeast extract..................................................................................1.0g
NH 4 Cl ...........................................................................................1.0g
KCl................................................................................................0.8g
Na 2 HPO 4 .......................................................................................0.6g
L- Cysteine·HCl·H 2 O....................................................................0.25g
Na 2 S·9H 2 O..................................................................................0.25g
CaCl 2 ·2H 2 O.................................................................................0.14g
Resazurin ...................................................................................1.0mg
Wolfe’s mineral solution..........................................................10.0mL
pH 7.2 ± 0.2 at 25°C
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.
Preparation of Medium: Add components, except Na 2 S·9H 2 O, to
glass-distilled water and bring volume to 990.0mL. Mix thoroughly.
Methanol or methylamine·HCl may be substituted for the trimethyl-
amine·HCl at a concentration of 50 mM. Heat gently and bring to boil-
ing. Adjust pH to 7.2 with 6N HCl. Autoclave for 5 min at 10 psi
pressure–115°C. Cool to 50°C under 80% N 2 + 20% CO 2 . If a large
precipitate is present, add a small amount of HCl and mix thoroughly.
Add Na 2 S·9H 2 O. Mix thoroughly. Distribute into tubes under 80% N 2
+ 20% CO 2 . Cap with butyl rubber stoppers. Autoclave for 15 min at
15 psi pressure–121°C. A precipitate will form but resolubilizes as the
medium cools. Invert tubes as they are cooling to facilitate resolubili-
zation. Allow tubes to cool in a slanted position.
Use:  For the cultivation and maintenance of Methanococcoides meth-
ylutens and Methanosarcina acetivorans.