Methanobacterium thermoautotrophicum 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 thermoautotrophicum 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:

Na 2 CO 3 .........................................................................................4.0g
(NH 4 ) 2 SO 4 ....................................................................................1.5g
NaCl..............................................................................................0.6g
KH 2 PO 4 .........................................................................................0.3g
K 2 HPO 4 .......................................................................................0.15g
MgSO 4 ·7H 2 O............................................................................. 0.12g
CaCl 2 ·2H 2 O ................................................................................0.08g
FeSO 4 ·7H 2 O.............................................................................. 4.0mg
Resazurin ...................................................................................1.0mg
Trace elements solution...........................................................10.0mL
Vitamin solution.......................................................................10.0mL
L -Cysteine·HCl solution ..........................................................10.0mL
Na 2 S·9H 2 O solution.................................................................10.0mL
pH 7.2 ± 0.2 at 25°C
Trace Elements Solution:
Composition per liter:
NaCl..............................................................................................1.0g
Disodium EDTA.........................................................................0.64g
MnSO 4 ·4H 2 O............................................................................. 0.55g
MgSO 4 ·7H 2 O............................................................................... 0.2g
ZnSO 4 ·7H 2 O.............................................................................. 0.18g
CoCl 2 ·6H 2 O................................................................................0.17g
CuSO 4 .........................................................................................0.15g
CaCl 2 ·2H 2 O ................................................................................0.13g
FeSO 4 ·7H 2 O................................................................................. 0.1g
NiCl 2 ·H 2 O.................................................................................0.025g
KAl(SO 4 ) 2 ·12H 2 O.....................................................................0.018g
Na 2 MoO 4 ·2H 2 O....................................................................... 0.011g
H 3 BO 3 .........................................................................................0.01g
Preparation of Trace Elements Solution : Add disodium EDTA
to 500.0mL of distilled/deionized water. Mix thoroughly to dissolve.
Add remaining components. Bring volume to 1.0L with distilled/de-
ionized water.
Vitamin Solution:
Composition per liter:
Pyridoxine·HCl........................................................................10.0mg
Calcium  DL -pantothenate...........................................................5.0mg
Nicotinic acid.............................................................................5.0mg
Riboflavin ..................................................................................5.0mg
Thiamine·HCl ............................................................................5.0mg
Biotin .........................................................................................2.0mg
Folic acid....................................................................................2.0mg
p-Aminobenzoic acid.................................................................1.0mg
Cyanocobalamin ......................................................................0.01mg
Preparation of Vitamin Solution:  Add components to distilled/
deionized water and bring volume to 1.0L. Adjust pH to 7.0. Mix thor-
oughly.
L -Cysteine·HCl Solution:
Composition per 10.0mL:
L -Cysteine·HCl..............................................................................1.5g
Preparation of  L -Cysteine·HCl Solution: Add  L -cysteine·HCl to
distilled/deionized water and bring volume to 10.0mL. Mix thorough-
ly. Autoclave under 80% N 2 + 20% CO 2 for 15 min at 15 psi pressure–
121°C.
Na 2 S·9H 2 O Solution:
Composition per 10.0mL:
Na 2 S·9H 2 O....................................................................................1.5g
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.
Sparge with 80% N 2 + 20% CO 2 . Autoclave for 15 min at 15 psi pres-
sure–121°C.
Preparation of Medium: Prepare and dispense medium under 80%
H 2 + 20% CO 2 . Add components, except  L -cysteine·HCl solution and
Na 2 S·9H 2 O solution, to distilled/deionized water and bring volume to
980.0mL. Mix thoroughly. Anaerobically distribute into tubes or flasks
fitted with butyl rubber stoppers. Autoclave for 15 min at 15 psi pres-
sure–121°C. Anaerobically add 10.0mL of sterile  L -cysteine·HCl solu-
tion and 10.0mL of sterile Na 2 S·9H 2 O solution to each liter of medium
or, using a syringe, inject the appropriate amount of sterile  L -
cysteine·HCl solution and sterile Na 2 S·9H 2 O solution into individual
tubes containing medium.
Use: For the cultivation and maintenance of Methanobacterium ther-
moautotrophicus and Methanoculleus oldenburgensis.