Halophile Medium III

[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]

Halophile Medium III

培养基配方

Composition per liter:
NaCl............................................................................................95.3g
MgSO 4 ·7H 2 O..............................................................................81.3g
Glycerol ........................................................................................5.0g
NH 4 Cl ...........................................................................................1.0g
K 2 HPO 4 solution...................................................................100.0mL
Wolfe's vitamin solution..........................................................10.0mL
Trace elements solution SL-10..................................................1.0mL
pH 7.5 ± 0.2 at 25°C
Wolfe's Vitamin Solution:
Composition per liter:
Pyridoxine·HCl........................................................................10.0mg
Calcium  D -(+)-pantothenate ......................................................5.0mg
Nicotinic acid.............................................................................5.0mg
p-Aminobenzoic acid.................................................................5.0mg
Riboflavin..................................................................................5.0mg
Thiamine·HCl............................................................................5.0mg
Thioctic acid..............................................................................5.0mg
Biotin.........................................................................................2.0mg
Folic acid ...................................................................................2.0mg
Cyanocobalamin.....................................................................100.0μg
Preparation of Wolfe’s Vitamin Solution: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Filter sterilize.
Trace Elements Solution SL-10:
Composition per liter:
FeCl 2 ·4H 2 O...................................................................................1.5g
CoCl 2 ·6H 2 O...........................................................................190.0mg
MnCl 2 ·4H 2 O ..........................................................................100.0mg
ZnCl 2 ........................................................................................70.0mg
Na 2 MoO 4 ·2H 2 O.......................................................................36.0mg
NiCl 2 ·6H 2 O..............................................................................24.0mg
H 3 BO 3 ........................................................................................6.0mg
CuCl 2 ·2H 2 O...............................................................................2.0mg
HCl (25% solution)..................................................................10.0mL
Preparation of Trace Elements Solution SL-10: Add FeCl 2 ·4H 2 O to 10.0mL of HCl solution. Mix thoroughly. Add distilled/deionized water and bring volume to 1.0L. Add remaining components. Mix thoroughly. Sparge with 100% N 2 . Autoclave for 15 min at 15 psi pressure–121°C.
K 2 HPO 4 Solution:
Composition per 100.0mL:
K 2 HPO 4 ........................................................................................1.0g
Preparation of  K 2 HPO 4 Solution: Add K 2 HPO 4 to distilled/deionized water and bring volume to 100.0mL. Mix thoroughly. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C.

Preparation of Medium: Add components, except K 2 HPO 4 solution, Wolfe’s vitamin solution, and trace elments solution SL-10, to distilled/deionized water and bring volume to 889.0mL. Mix thoroughly. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C. Aseptically add 100.0mL of sterile K 2 HPO 4 solution, 10.0mL of sterile Wolfe's vitamin solution, and 1.0mL of sterile trace elements solution SL-10. Mix thoroughly. Aseptically adjust pH to 7.5. Aseptically distribute into sterile tubes or flasks.

Use: For the cultivation and maintenance of Halovibrio variabilis.