Haloanaerobium congolense Medium (DSMZ Medium 933)

[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 congolense Medium

(DSMZ Medium 933)

培养基配方

Composition per 1080.0mL:
NaCl..........................................................................................100.0g
MgCl 2 ·6H 2 O................................................................................10.0g
Trypticase™..................................................................................1.0g
NH 4 Cl ...........................................................................................1.0g
KCl............................................................................................... 1.0g
Na-acetate .....................................................................................0.5g
Cysteine ........................................................................................0.5g
K 2 HPO 4 .........................................................................................0.3g
KH 2 PO 4 .........................................................................................0.3g
CaCl 2 ·2H 2 O...................................................................................0.1g
Resazurin ....................................................................................0.01g
Glucose solution ......................................................................20.0mL
Thiosulfate solution .................................................................20.0mL
Na 2 S·9H 2 O solution.................................................................20.0mL
NaHCO 3  solution .....................................................................20.0mL
Trace elements solution .............................................................1.0mL

pH 7.0 ± 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....................................................................................0.2g
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. Sparge with 100% N 2 . Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C.
NaHCO 3 Solution:
Composition per 50.0mL:
NaHCO 3 ........................................................................................5.0g
Preparation of NaHCO 3 Solution:  Add NaHCO 3 to distilled/deionized water and bring volume to 50.0mL. Mix thoroughly. Sparge with 80% N 2 + 20% CO 2 . Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C.
Thiosulfate Solution:
Composition per 20.0mL:
Na 2 S 2 O 3 ·5H 2 O..............................................................................5.0g
Preparation of Thiosulfate Solution:  Add Na 2 S 2 O 3 ·5H 2 O 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.
Cool to room temperature.
Glucose Solution:
Composition per 20.0mL:
Glucose.........................................................................................3.5g
Preparation of Glucose Solution: Add glucose to distilled/deionized water and bring volume to 20.0mL. Mix thoroughly. Sparge with 100% N 2 . Filter sterilize.
Preparation of Medium: Prepare and dispense medium under 80% N 2 + 20% CO 2 gas atmosphere. Add components, except NaHCO 3  solution, glucose solution, Na 2 S·9H 2 O solution, and thiosulfate solution, to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently heat and bring to boiling. Boil for 5 min. Cool to room temperature while sparging with 80% N 2 + 20% CO 2 . Adjust pH to 7.0. Distribute into anaerobe tubes or bottles. Autoclave for 15 min at 15 psi pressure–121°C. Cool to room temperature. Aseptically and anaerobically add per 10.0mL medium, 0.2mL NaHCO 3  solution, 0.2mL glucose solution, 0.2mL Na 2 S·9H 2 O solution, and 0.2mL thiosulfate solution. Mix thoroughly. The final pH should be 7.0.
Use:  For the cultivation of Thermococcus waiotapuensis.