Methanolobus taylorii Medium (DSMZ Medium 490a)

[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 taylorii Medium
(DSMZ Medium 490a)

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............................................................................................29.0g
Yeast extract..................................................................................2.0g
Trypticase™ peptone....................................................................2.0g
MgCl 2 ·6H 2 O .................................................................................1.7g
KCl................................................................................................1.5g
NH 4 Cl ...........................................................................................1.0g
Resazurin ......................................................................................1.0g
Mercaptoethanesulfonic acid (coenzyme M)................................0.5g
K 2 HPO 4 ·3H 2 O ..............................................................................0.4g
Trace elements solution...........................................................10.0mL
Trimethylamine·HCl solution..................................................10.0mL
Na 2 S·9H 2 O solution.................................................................10.0mL
NaHCO 3 solution.....................................................................10.0mL
pH 8.5 ± 0.2 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
H 3 BO 3 .........................................................................................0.01g
Na 2 MoO 4 ·4H 2 O..........................................................................0.01g
CuSO 4 ·5H 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. Dissolve by adjusting pH
to 6.5 with KOH. Add remaining components. Add distilled/deionized
water to 1.0L. Mix thoroughly.
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. Sparge with N 2 .
Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C. Store an-
aerobically.
Trimethylamine·HCl Solution:
Composition per 10.0mL:
Trimethylamine·HCl.....................................................................2.0g
Preparation of Trimethylamine·HCl Solution:  Add trimethyl-
amine·HCl to distilled/deionized water and bring volume to 10.0mL.
Mix thoroughly. Autoclave for 15 min at 15 psi pressure–121°C. Cool
to 25°C.
NaHCO 3 Solution:
Composition per 10.0mL:
NaHCO 3 ........................................................................................2.0g
Preparation of NaHCO 3 Solution:  Add NaHCO 3 to distilled/de-
ionized water and bring volume to 10.0mL. Mix thoroughly. Autoclave
for 15 min at 15 psi pressure–121°C. Cool to 25°C. Should be prepared
freshly.
Preparation of Medium: Prepare and dispense medium under an
oxygen-free 100% N 2 gas mixture. Add components, except
trimethylamine·HCl solution, NaHCO 3 solution, and Na 2 S·9H 2 O solu-
tion, to 970.0mL distilled/deionized water. Mix thoroughly. Sparge for
20 min with 100% N 2 . Adjust pH to 8.0. Autoclave for 15 min at 15 psi
pressure–121°C. Cool to 25°C. Aseptically and anaerobically add
10.0mL sterile trimethylamine·HCl solution, and 10.0mL Na 2 S·9H 2 O
solution. Adjust pH to 8.5 using sterile NaHCO 3 solution, approxi-
mately 10.0mL per liter medium. Aseptically and anaerobically dis-
tribute to sterile tubes or flasks under 100% N 2 .
Use:  For the cultivation of Methanolobus taylorii.