Haloanaerobium alcaliphilum Medium (DSMZ Medium 807)

[所属分类:培养基配方] [发布时间:2021-3-24] [发布人: ] [阅读次数:] [返回]

Haloanaerobium alcaliphilum Medium

(DSMZ Medium 807)

培养基配方

Composition per liter:
NaCl..........................................................................................100.0g
MgSO 4 ·7H 2 O..............................................................................17.0g
Trypticase™................................................................................10.0g
NaHCO 3 ........................................................................................4.1g
Cysteine-HCl·H 2 O........................................................................0.5g
Resazurin ...................................................................................1.0mg
Solution A................................................................................50.0mL
Solution B................................................................................50.0mL
Yeast extract solution...............................................................50.0mL
Glucose solution ......................................................................20.0mL
Trace elements solution...........................................................10.0mL
Na 2 S·9H 2 O solution.................................................................10.0mL
pH 7.0 ± 0.1 at 25°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 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.
Glucose Solution :
Composition per 20.0mL:
Glucose.........................................................................................5.0g
Preparation of Glucose Solution:  Add glucose to distilled/deionized water and bring volume to 20.0mL. Mix thoroughly. Filter sterilize.
Yeast Extract Solution :
Composition per 50.0mL:
Yeast extract................................................................................10.0g
Preparation of Yeast Extract Solution:  Add yeast extract to distilled/deionized water and bring volume to 50.0mL. Mix thoroughly. Autoclave under 100% N 2 for 15 min at 15 psi pressure–121°C. Cool to room temperature.
Solution A
Composition per liter:
K 2 HPO 4 .........................................................................................6.0g
Preparation of Solution A: Add K 2 HPO 4 to distilled/deionized water and bring volume to 1.0L. Mix thoroughly.
Solution B
Composition per liter:
NaCl............................................................................................12.0g
KH 2 PO 4 .........................................................................................6.0g
(NH 4 )SO 4 ......................................................................................6.0g
MgSO 4 ·7H 2 O................................................................................2.6g

NH 4 Cl ...........................................................................................2.5g

CaCl 2 ·2H 2 O.................................................................................0.28g
K 2 HPO 4 .......................................................................................0.28g
Preparation of Solution B: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly.
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.
Preparation of Medium: Prepare and dispense medium under 80% N 2 + 20% CO 2 gas atmosphere. Add components, except NaHCO 3 , Na 2 S·9H 2 O solution, cysteine-HCl·H 2 O, yeast extract solution, and glucose 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 under 80% N 2 + 20% CO 2 gas atmosphere. Add 4.1g NaHCO 3  and 0.5g cysteine-HCl·H 2 O. Adjust pH to 7.0. Autoclave for 15 min at 15 psi pressure–121°C. Aseptically and anaerobically add 20.0mL sterile glucose solution, 50.0mL sterile yeast extract solution, and 10.0mL sterile Na 2 S·9H 2 O solution. Mix thoroughly. Aseptically and anaerobically distribute into sterile tubes or bottles.
Use:  For the cultivation of Halanaerobium alcaliphilum.