Methanospirillum hungatei 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]

Methanospirillum hungatei Medium

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

Composition per 100.0mL:

Na 2 CO 3 .........................................................................................0.4g
Sodium formate.............................................................................0.2g
Pancreatic digest of casein............................................................0.2g
Yeast extract..................................................................................0.2g
NaCl............................................................................................0.05g
L- Cysteine·HCl·H 2 O....................................................................0.03g
K 2 HPO 4 .......................................................................................0.02g
KH 2 PO 4 .......................................................................................0.02g
(NH 4 ) 2 SO 4 ...................................................................................0.02g
MgSO 4 ·7H 2 O.............................................................................9.0mg
CaCl 2 ·2H 2 O................................................................................6.0mg
Resazurin ...................................................................................0.1mg
Na 2 S·9H 2 O solution.................................................................10.0mL
Vitamin solution.........................................................................1.0mL
Trace metal solution...................................................................1.0mL
pH 7.0 ± 0.2 at 25°C
Na 2 S·9H 2 O Solution:
Composition per 10.0.mL:
Na 2 S·9H 2 O..................................................................................0.03g
Preparation of Na 2 S·9H 2 O Solution: Add Na 2 S·9H 2 O to dis-
tilled/deionized water and bring volume to 10.0mL. Mix thoroughly.
Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C.
Vitamin Solution:
Composition per 1000.0mL:
Pyridoxine·HCl..........................................................................1.0mg
p-Aminobenzoic acid.................................................................0.5mg
Calcium- D- pantothenate.............................................................0.5mg
Nicotinic acid.............................................................................0.5mg
Riboflavin ..................................................................................0.5mg
Thiamine·HCl ............................................................................0.5mg
Thioctic acid ..............................................................................0.5mg
Biotin .........................................................................................0.2mg
Folic acid....................................................................................0.2mg
Vitamin B 12 ..............................................................................0.01mg
Preparation of Vitamin Solution: Add components to distilled/
deionized water and bring volume to 1.0L. Mix thoroughly. Filter ster-
ilize.
Trace Metal Solution:
Composition per liter:
K 2 HPO 4 ·3H 2 O ..............................................................................9.0g
K 2 HPO 4 .........................................................................................6.0g
NH 4 Cl ...........................................................................................5.0g
MgCl 2 ·6H 2 O .................................................................................1.0g
CaCl 2 ·2H 2 O ................................................................................0.01g
Preparation of Trace Metal Solution: Add components to dis-
tilled/deionized water and bring volume to 1.0L. Mix thoroughly.
Preparation of Medium: Prepare and distribute medium anaerobi-
cally under 80% H 2 + 20% CO 2 . Add components, except Na 2 S·9H 2 O
solution, to distilled/deionized water and bring volume to 90.0mL. Mix
thoroughly. Gently heat and bring to boiling. Continue boiling until re-
sazurin turns colorless, indicating reduction. Autoclave for 15 min at
15 psi pressure–121°C. Cool to 25°C. Aseptically add 10.0mL of ster-
ile Na 2 S·9H 2 O solution. Mix thoroughly. Aseptically and anaerobical-
ly distribute into sterile tubes or flasks.
Use: For the cultivation of Methanospirillum hungatei.