Halorhodospira Medium (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

(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
CaCl 2 ·2H 2 O.................................................................................0.05g
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 SL12:  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. Aseptically add sulfide, bicarbonate, vitamin, and trace elements solutions. Mix thoroughly. Distribute into screw-cap tubes, bottles, or flasks. Autoclave for 20 min at 15 psi pressure–121°C.
Use: For the cultivation of Halorhodospira spp.