Methanogenium CV Medium

[所属分类:培养基配方] [发布时间:2021-5-25] [发布人:网站管理员2] [阅读次数:] [返回]

Methanogenium CV 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
Isopropanol ...................................................................................7.5g
NaHCO 3 ........................................................................................5.0g
MgCl 2 ·6H 2 O..................................................................................4.0g
MgSO 4 ·7H 2 O............................................................................. 3.45g
Trypticase™..................................................................................2.0g
Yeast extract..................................................................................2.0g
Sodium acetate..............................................................................1.0g
KCl............................................................................................0.335g
NH 4 Cl .........................................................................................0.25g
CaCl 2 ·2H 2 O.................................................................................0.14g
K 2 HPO 4 .......................................................................................0.14g
L -Cysteine·HCl..............................................................................0.5g
Na 2 S·9H 2 O....................................................................................0.4g
Fe(NH 4 ) 2 (SO 4 ) 2 ·7H 2 O ...............................................................2.0mg
Resazurin ...................................................................................1.0mg
Na 2 WoO 4 ·2H 2 O .......................................................................0.03mg
Trace elements solution ...........................................................10.0mL
Vitamin solution.......................................................................10.0mL
pH 6.8–7.0 at 25°C
Trace Elements Solution:
Composition per liter:
MgSO 4 ·7H 2 O............................................................................... 3.0g
Nitrilotriacetic acid.......................................................................1.5g
NaCl..............................................................................................1.0g
MnSO 4 ·2H 2 O............................................................................... 0.5g
CoSO 4 ·7H 2 O...............................................................................0.18g
ZnSO 4 ·7H 2 O...............................................................................0.18g
CaCl 2 ·2H 2 O ..................................................................................0.1g
FeSO 4 ·7H 2 O..................................................................................0.1g
NiCl 2 ·6H 2 O...............................................................................0.025g
KAl(SO 4 ) 2 ·12H 2 O.......................................................................0.02g
CuSO 4 ·5H 2 O...............................................................................0.01g
H 3 BO 3 .........................................................................................0.01g
Na 2 MoO 4 ·2H 2 O......................................................................... 0.01g
Na 2 SeO 3 ·5H 2 O...........................................................................0.3mg
Preparation of Trace Elements Solution: Add nitrilotriacetic
acid to 500.0mL of distilled/deionized water. Adjust pH to 6.5 with
KOH. Add remaining components. Adjust pH to 7.0. Add distilled/de-
ionized water to 1.0L.
Vitamin Solution:
Composition per liter:
Pyridoxine·HCl........................................................................10.0mg
Calcium  DL -pantothenate...........................................................5.0mg
Nicotinic acid.............................................................................5.0mg
p-Aminobenzoic acid.................................................................5.0mg
Lipoic acid.................................................................................5.0mg
Riboflavin..................................................................................5.0mg
Thiamine·HCl............................................................................5.0mg
Biotin.........................................................................................2.0mg
Folic acid ...................................................................................2.0mg
Vitamin B 12 ................................................................................0.1mg
Preparation of Vitamin Solution : Add components to distilled/
deionized water and bring volume to 1.0L. Mix thoroughly. Filter ster-
ilize. Sparge with 80% H 2 + 20% CO 2 .
Preparation of Medium: Prepare and dispense medium under 80%
H 2 + 20% CO 2 . Add components, except vitamin solution, to distilled/
deionized water and bring volume to 990.0mL. Mix thoroughly. Adjust
pH to 6.8–7.0. Anaerobically distribute into tubes or flasks fitted with
butyl rubber stoppers. Autoclave for 15 min at 15 psi pressure–121°C.
Anaerobically add 10.0mL of sterile vitamin solution to each liter of
medium or, using a syringe, inject the appropriate amount of sterile vi-
tamin solution into individual tubes containing medium.
Use:  For the cultivation and maintenance of Methanogenium organo-
philum.