Halomonas subglaciescola Medium (Artificial Organic Lake Medium)

[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]

Halomonas subglaciescola Medium

(Artificial Organic Lake Medium)

培养基配方

Composition per 1001.0mL:
NaCl............................................................................................80.0g
MgSO 4 ·7H 2 O................................................................................9.5g
Yeast extract..................................................................................1.0g
KCl................................................................................................0.5g
CaCl 2 ·2H 2 O ..................................................................................0.2g
KNO 3 ............................................................................................0.1g
(NH 4 ) 2 SO 4 ....................................................................................0.1g
Hutner's basal salts solution.....................................................20.0mL
Phosphate supplement .............................................................20.0mL
Vitamin solution.........................................................................1.0mL
pH 8.0 ± 0.2 at 25°C
Hutner’s Basal Salts Solution:
Composition per liter:
MgSO 4 ·7H 2 O..............................................................................29.7g
Nitrilotriacetic acid.....................................................................10.0g
CaCl 2 ·2H 2 O ..............................................................................3.335g
FeSO 4 ·7H 2 O.............................................................................99.0mg
(NH 4 ) 6 Mo 7 O 24 ·4H 2 O...............................................................9.25mg
“Metals 44”..............................................................................50.0mL
“Metals 44”:
Composition per 100.0mL:
ZnSO 4 ·7H 2 O.............................................................................1.095g
FeSO 4 ·7H 2 O..................................................................................0.5g
Sodium EDTA.............................................................................0.25g
MnSO 4 ·H2O .............................................................................0.154g
CuSO 4 ·5H 2 O............................................................................39.2mg
Co(NO 3 ) 2 ·6H 2 O.......................................................................24.8mg
Na 2 B 4 O 7 ·10H 2 O.......................................................................17.7mg
Preparation of “Metals 44”: Add sodium EDTA to distilled/deionized water and bring volume to 90.0mL. Mix thoroughly. Add a few drops of concentrated H 2 SO 4 to retard precipitation of heavy metalions. Add remaining components. Mix thoroughly. Bring volume to 100.0mL with distilled/deionized water.
Preparation of Hutner’s Basal Salts Solution:  Add nitrilotriacetic acid to 500.0mL of distilled/deionized water. Adjust pH to 6.5 with KOH. Add remaining components. Add distilled/deionized water to 1.0L. Adjust pH to 6.8. Filter sterilize.
Phosphate Supplement:
Composition per 20.0mL:
K 2 HPO 4 ....................................................................................50.0mg
KH 2 PO 4 ....................................................................................50.0mg
Preparation of Phosphate Supplement:  Add components to distilled/deionized water and bring volume to 20.0mL. Mix thoroughly. Filter sterilize.
Vitamin Solution:
Composition per liter:
Cyanocobalamin......................................................................10.0mg
Pyridoxine·HCl........................................................................10.0mg
Thiamine·HCl..........................................................................10.0mg
Calcium  DL -pantothenate...........................................................5.0mg
Nicotinamide..............................................................................5.0mg
Biotin .........................................................................................2.0mg
Folic acid....................................................................................2.0mg
Preparation of Vitamin Solution:  Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Filter sterilize.

Preparation of Medium: Add components, except Hutner's basal salts solution, phosphate supplement, and vitamin solution, to distilled/deionized water and bring volume to 960.0mL. Mix thoroughly. Adjust pH to 8.0. Autoclave for 15 min at 15 psi pressure–121°C. Aseptically add 20.0mL of sterile Hutner's basal salts solution, 20.0mL of sterile phosphate supplement, and 1.0mL of sterile vitamin solution. Mix thoroughly. Aseptically distribute into sterile tubes or flasks.

Use: For the cultivation of Halomonas subglaciescola.