Methanohalophilus oregonense 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]

Methanohalophilus oregonense Medium

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

Composition per 1010.0mL:

NaCl............................................................................................29.0g
Trimethylamine·HCl.....................................................................2.0g
Trypticase™..................................................................................2.0g
Yeast extract..................................................................................2.0g
MgCl 2 ·6H 2 O..................................................................................1.7g
KCl................................................................................................1.5g
NH 4 Cl ...........................................................................................1.0g
Resazurin ......................................................................................1.0g
Coenzyme M (mercaptoethane
sulfonic acid)..........................................................................0.5g
K 2 HPO 4 ·3H 2 O...............................................................................0.4g
Na 2 S·9H 2 O solution.................................................................10.0mL
Trace elements solution ...........................................................10.0mL
Na 2 CO 3 solution......................................................................variable
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
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
NiCl 2 ·6H 2 O...............................................................................0.025g
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.
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.
Sparge with 100% N 2 . Autoclave for 15 min at 15 psi pressure–121°C.
Na 2 CO 3 Solution:
Composition per 10.0mL:
Na 2 CO 3 .........................................................................................1.0g
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. Sparge with
100% N 2 . Autoclave for 15 min at 15 psi pressure–121°C.
Preparation of Medium: Prepare and dispense medium under 80%
N 2 + 20% CO 2 . Add components, except Na 2 S·9H 2 O solution, to dis-
tilled/deionized water and bring volume to 1.0L. Mix thoroughly.
Sparge with 80% N 2 + 20% CO 2 . Autoclave for 15 min at 15 psi pres-
sure–121°C. Aseptically and anaerobically add 10.0mL of sterile
Na 2 S·9H 2 O solution. Mix thoroughly. Adjust pH to 8.5 with a suffi-
cient quantity of sterile Na 2 CO 3 solution (approximately 0.1mL per
10.0mL of medium).
Use: For the cultivation and maintenance of Methanohalophilus ore-
gonense.