Halorhodospira Medium with Succinate, Acetate, and Yeast Extract (DSMZ Medium 999)

[所属分类:培养基配方] [发布时间:2021-3-30] [发布人: ] [阅读次数:] [返回]

Halorhodospira Medium with Succinate, Acetate, and Yeast Extract

(DSMZ Medium 999)

培养基配方

Composition per liter:
NaCl..........................................................................................100.0g
MgCl 2 ·6H 2 O .................................................................................2.0g
MgSO 4 · 7H 2 O................................................................................1.0g
KH 2 PO 4 ........................................................................................1.0g
NH 4 Cl ..........................................................................................0.5g
Yeast extract..................................................................................0.5g
CaCl 2 ·2H 2 O ................................................................................0.05g
Na-succinate ...............................................................................0.27g
Na-acetate...................................................................................0.17g
NaHCO 3  solution.....................................................................10.0mL
Na 2 S·9H 2 O solution...................................................................7.5mL
Vitamin solution V7 ..................................................................1.0mL
Trace elements solution SL-12..................................................1.0mL
pH 7.3 ± 0.2 at 25°C
Na 2 S·9H 2 O Solution :
Composition per 10.0mL:
Na 2 S·9H 2 O....................................................................................1.0g
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.
NaHCO 3 Solution :
Composition per 10.0mL:
NaHCO 3 ........................................................................................2.0g
Preparation of NaHCO 3 Solution:  Add components to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Sparge with 20% CO 2 + 80% H 2 . Autoclave for 15 min at 15 psi pressure–121°C. Cool to room temperature.
Vitamin Solution V7:
Composition per liter:
Pyridoxine-HCl........................................................................50.0mg
Nicotinic acid...........................................................................20.0mg
Vitamin B 12 ..............................................................................20.0mg
Thiamine-HCl·2H 2 O................................................................10.0mg
p-Aminobenzoic acid...............................................................10.0mg
D -Ca-pantothenate......................................................................5.0mg
Biotin .........................................................................................2.0mg
Preparation of Vitamin Solution V7:  Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Filter sterilize.
Trace Elements Solution SL-12:
Composition per liter:
FeSO 4 ·7H 2 O..................................................................................1.1g
H 3 BO 3 ...........................................................................................0.3g
CoCl 2 ·6H 2 O ................................................................................0.19g
MnCl 2 ·2H 2 O................................................................................0.05g
ZnCl 2 ........................................................................................42.0mg
NiCl 2 ·6H 2 O..............................................................................24.0mg
Na 2 MoO 4 ·4H 2 O .......................................................................18.0mg
CuCl 2 ·2H 2 O ...............................................................................2.0mg
Preparation of Trace Elements Solution SL-12:  Add components. to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Filter sterilize.
Preparation of Medium: Add components, except sulfide, bicarbonate, vitamin, and trace elements solutions, to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Adjust pH to 7.3. Gently heat while stirring and bring to boiling. Autoclave for 20 min at 15 psi pressure–121°C. Cool to room temperature. Aseptically add sulfide, bicarbonate, vitamin, and trace elements solutions. Mix thoroughly. Aseptically distribute into sterile screw-cap tubes, bottles, or flasks.
Use: For the improved growth during cultivation of pure cultures of Halorhodospira spp.