Methanolobus 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]

Methanolobus 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............................................................................................18.0g
NaHCO 3 ........................................................................................5.0g
MgSO 4 · 7H 2 O..............................................................................3.45g
MgCl 2 ·6H 2 O ...............................................................................2.75g
L- Cysteine·HCl·H 2 O .....................................................................0.5g
Na 2 S·9H 2 O....................................................................................0.5g
KCl............................................................................................0.335g
NH 4 Cl .........................................................................................0.25g
CaCl 2 ·2H 2 O ................................................................................0.14g
K 2 HPO 4 .......................................................................................0.14g
Fe(NH 4 ) 2 (SO 4 ) 2 ·6H 2 O...............................................................2.0mg
Resazurin...................................................................................1.0mg
Wolfe’s mineral solution..........................................................10.0mL
Wolfe’s vitamin solution..........................................................10.0mL
Methanol....................................................................................5.0mL
pH 6.5 ± 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.
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.
Filter sterilize.
Preparation of Medium: Prepare and dispense medium under 80%
N 2 + 20% CO 2 . Add components, except methanol and Wolfe’s vitamin
solution, to distilled/deionized water and bring volume to 985.0mL. Mix
thoroughly. Autoclave for 15 min at 15 psi pressure–121°C. Cool under
80% N 2 + 20% CO 2 . Aseptically add sterile Wolfe’s vitamin solution and
sterile methanol. Adjust pH to 6.5. Aseptically and anaerobically distrib-
ute into sterile tubes or flasks.
Use: For the cultivation and maintenance of Methanolobus siciliae
and Methanolobus tindarius.