Methanohalophilus zhilinae 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 zhilinae 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............................................................................................40.0g
MgCl 2 ·6H 2 O .................................................................................3.5g
MgSO 4 ·7H 2 O................................................................................3.0g
Trypticase™..................................................................................2.0g
Yeast extract..................................................................................2.0g
KCl................................................................................................1.0g
NH 4 Cl ...........................................................................................1.0g
L -Cysteine·HCl .............................................................................0.5g
K 2 HPO 4 .........................................................................................0.4g
Resazurin...................................................................................1.0mg
Na 2 SeO 3 ·5H 2 O...........................................................................0.1mg
Trimethylamine·HCl solution..................................................20.0mL
NaHCO 3  solution.....................................................................10.0mL
Na 2 CO 3  solution.......................................................................10.0mL
Na 2 S·9H 2 O solution.................................................................10.0mL
Trace elements solution.............................................................5.0mL
pH 9.2 ± 0.2 at 25°C
Trimethylamine·HCl Solution:
Composition per 20.0mL:
Trimethylamine·HCl.....................................................................2.0g
Preparation of Trimethylamine·HCl Solution:  Add trimethyl-
amine·HCl to distilled/deionized water and bring volume to 20.0mL.
Mix thoroughly. Sparge with 100% N 2 . Autoclave for 15 min at 15 psi

pressure–121°C.

NaHCO 3 Solution:
Composition per 10.0mL:
NaHCO 3 ........................................................................................0.5g
Preparation of NaHCO 3 Solution:  Add NaHCO 3 to distilled/de-
ionized 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 .........................................................................................2.0g
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.
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.
Sparge with 100% N 2 . Autoclave for 15 min at 15 psi pressure–121°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.
Preparation of Medium: Prepare and dispense medium under
100% N 2 . Add components, except trimethylamine·HCl solution,
NaHCO 3  solution, Na 2 CO 3 solution,  L -cysteine·HCl, and Na 2 S·9H 2 O
solution, to distilled/deionized water and bring volume to 950.0mL.
Mix thoroughly. Gently heat and bring to boiling. Continue boiling for
5 min. Allow to cool to room temperature while sparging with 100%
N 2 . Add  L -cysteine·HCl. Distribute medium into tubes and seal. Auto-
clave for 15 min at 15 psi pressure–121°C. Anaerobically add 20.0mL
of sterile trimethylamine·HCl solution, 10.0mL of sterile NaHCO 3  so-
lution, 10.0mL of sterile Na 2 CO 3 solution, and 10.0mL of sterile
Na 2 S·9H 2 O solution. Mix thoroughly. Check pH.
Use:  For the cultivation and maintenance of Methanohalophilus zhili-
nae.