Methanococcus vannielii Medium

[所属分类:培养基配方] [发布时间:2021-5-24] [发布人:网站管理员2] [阅读次数:] [返回]

Methanococcus vannielii Medium

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

Composition per 1020.0mL:

Solution A..............................................................................500.0mL
Inorganic salts solution..........................................................500.0mL
Na 2 S·9H 2 O solution.................................................................10.0mL
Na 2 CO 3 solution.......................................................................10.0mL
Solution A:
Composition per 500.0mL:
Sodium formate ..........................................................................10.0g
Phenol Red.................................................................................3.0mg
Methylene Blue..........................................................................2.0mg
Preparation of Solution A: Add components to distilled/deionized
water and bring volume to 500.0mL. Mix thoroughly. Autoclave for 15
min at 15 psi pressure–121°C. Cool to 25°C.
Inorganic Salts Solution:
Composition per 500.0mL:
K 2 HPO 4 ·3H 2 O ............................................................................1.45g
NH 4 Cl ...........................................................................................1.0g
KH 2 PO 4 .......................................................................................0.75g
MgCl 2 ·6H 2 O .................................................................................0.2g
Nitrilotriacetic acid.....................................................................0.04g
CaCl 2 ·2H 2 O ................................................................................0.02g
FeCl 2 ·4H 2 O................................................................................3.6mg
CoCl 2 ·6H 2 O...............................................................................1.5mg
MnCl 2 ·4H2O..............................................................................0.9mg
ZnCl 2 ..........................................................................................0.9mg
H 3 BO 2 ......................................................................................0.17mg
Na 2 MoO 4 ·2H 2 O.......................................................................0.09mg
Preparation of Inorganic Salts Solution: Add nitrilotriacetic
acid to 250.0mL of distilled/deionized water. Dissolve by adjusting pH
to 6.5 with KOH. Add remaining components. Readjust pH to 7.2 with
H 2 SO 4 or KOH. Add distilled/deionized water to 500.0mL. Filter ster-
ilize.
Na 2 S·9H 2 O Solution:
Composition per 10.0mL:
Na 2 S·9H 2 O....................................................................................0.3g
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.
Na 2 CO 3 Solution:
Composition per 10.0mL:
Na 2 CO 3 .........................................................................................2.5g
Preparation of Na 2 CO 3 Solution: Add Na 2 CO 3 to distilled/deion-
ized water and bring volume to 10.0mL. Mix thoroughly. Autoclave for
15 min at 15 psi pressure–121°C. Cool to 25°C.
Preparation of Medium: Prepare and distribute medium anaerobi-
cally under 80% N 2 + 20% CO 2 . Aseptically and anaerobically com-
bine 500.0mL of sterile inorganic salts solution, 500.0mL of sterile
solution A, 10.0mL of sterile Na 2 S·9H 2 O solution, and 10.0mL of ster-
ile Na 2 CO 3 solution. Mix thoroughly. Aseptically and anaerobically
distribute into sterile tubes or flasks.
Use: For the isolation and cultivation of Methanococcus vannielii
from marine mud.