Halonatronum saccharophilum Medium (DSMZ Medium 932)

[所属分类:培养基配方] [发布时间:2021-3-29] [发布人: ] [阅读次数:] [返回]

Halonatronum saccharophilum Medium

(DSMZ Medium 932)

培养基配方

Composition per liter:
Na 2 CO 3 .......................................................................................68.0g
NaCl............................................................................................50.0g
NaHCO 3 ......................................................................................38.0g
NH 4 Cl ...........................................................................................0.5g
KCl................................................................................................0.2g
KH 2 PO 4 .........................................................................................0.2g
MgCl 2 ............................................................................................0.1g
Resazurin ....................................................................................0.01g
Sucrose solution.......................................................................50.0mL
Na 2 S·9H 2 O solution.................................................................10.0mL
Yeast extract solution...............................................................10.0mL
Vitamin solution.......................................................................10.0mL
Trace elements...........................................................................1.0mL
pH 9.5–10.0 at 25°C
Sucrose Solution:
Composition per 50.0mL:
Sucrose..........................................................................................5.0g
Preparation of Sucrose Solution: Add sucrose to distilled/deionized water and bring volume to 50.0mL. Mix thoroughly. Sparge with 100% N 2 . Autoclave for 15 min at 15 psi pressure–121°C. Cool to room temperature.
Yeast Extract Solution:
Composition per 10.0mL:
Yeast extract..................................................................................0.2g
Preparation of Yeast Extract Solution: Add yeast extract to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Sparge with 100% N 2 . Autoclave under 100% N 2 for 15 min at 15 psi pressure–121°C. Cool to room temperature.
Na 2 S·9H 2 O Solution :
Composition per 10.0mL:
Na 2 S·9H 2 O....................................................................................0.7g
Preparation of Na 2 S·9H 2 O Solution: Add Na 2 S·9H 2 O to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Autoclave under 100% N 2 for 15 min at 15 psi pressure–121°C. Cool to room temperature.
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.
Vitamin Solution:
Composition per liter:
Pyridoxine-HCl........................................................................10.0mg
Thiamine-HCl·2H 2 O..................................................................5.0mg
Riboflavin..................................................................................5.0mg
Nicotinic acid.............................................................................5.0mg
D -Ca-pantothenate .....................................................................5.0mg
p-Aminobenzoic acid.................................................................5.0mg
Lipoic acid.................................................................................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. Sparge with 80% H 2 + 20% CO 2 . Filter sterilize.
Preparation of Medium: Prepare and dispense medium under 100% N 2 gas atmosphere. Add components, except NaHCO 3 , NH 4 Cl, Na 2 CO 3 , sucrose solution, Na 2 S·9H 2 O solution, yeast extract solution, and vitamin solution, to distilled/deionized water and bring volume to 920.0mL. Mix thoroughly. Gently heat and bring to boiling. Boil for 5 min. Cool to room temperature while sparging with 100% N 2 . Add solid NaHCO 3 , NH 4 Cl, and Na 2 CO 3 . Mix thoroughly. Distribute into anaerobe tubes or bottles. Autoclave for 15 min at 15 psi pressure–121°C. Aseptically and anaerobically add per liter of medium 50.0mL sucrose solution, 10.0mL yeast extract solution, 10.0mL Na 2 S·9H 2 O solution, and 10.0mL vitamin solution. The final pH should be 9.5–10.0.
Use:  For the cultivation of Halonatronum saccharophilum.