Methanobacterium ruminantium 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]

Methanobacterium ruminantium 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 ........................................................................................6.0g
NaCl..............................................................................................2.0g
L -Cysteine·HCl·H 2 O.....................................................................1.0g
K 2 HPO 4 ·3H 2 O ..............................................................................1.0g
KH 2 PO 4 .........................................................................................1.0g
NH 4 Cl ...........................................................................................1.0g
CaCl 2 ·2H 2 O ..................................................................................0.1g
MgSO 4 ·7H 2 O................................................................................0.1g
Resazurin ...................................................................................1.0mg
Rumen fluid...........................................................................300.0mL
Na 2 S·9H 2 O solution.................................................................10.0mL
6.8 ± 0.2 at 25°C
Na 2 S·9H 2 O Solution:
Composition per 10.0mL:
Na 2 S·9H 2 O..................................................................................0.25g
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.
Preparation of Medium: Prepare and distribute medium anaerobi-
cally under 80% H 2 + 20% CO 2 . Add components, except rumen fluid
and Na 2 S·9H 2 O solution, to distilled/deionized water and bring volume
to 690.0mL. Mix thoroughly. Gently heat and bring to boiling. Contin-
ue boiling until resazurin turns colorless, indicating reduction. Auto-
clave for 15 min at 15 psi pressure–121°C. Cool to 25°C. Aseptically
add 10.0mL of sterile Na 2 S·9H 2 O solution and 300.0mL of sterile ru-
men fluid. Mix thoroughly. Aseptically and anaerobically distribute
into sterile tubes or flasks.
Use: For the cultivation of Methanobacterium ruminantium.