Geoalkalibacter Medium (DSMZ Medium 1059)

[所属分类:培养基配方] [发布时间:2021-3-10] [发布人: ] [阅读次数:] [返回]

Geoalkalibacter Medium

(DSMZ Medium 1059)

培养基配方

Composition per liter:
Sulfur, powdered.........................................................................10.0g
NaHCO 3 ......................................................................................10.0g
Na 2 CO 3 .........................................................................................3.0g
NaCl..............................................................................................1.0g
NH 4 Cl ..........................................................................................0.5g
KCl................................................................................................0.2g
KH 2 PO 4 .........................................................................................0.2g
MgCl 2 · 6H 2 O..................................................................................0.1g
Yeast extract..................................................................................0.1g
Resazurin ...................................................................................0.5mg
Sodium acetate solution...........................................................10.0mL
Trace elements solution SL-10 ..................................................1.0mL
Selenite/tungstate solution.........................................................1.0mL
pH 9.1 ± 0.2 at 25°C
Sodium Acetate Solution:
Composition per 10.0mL:
Sodium acetate..............................................................................2.5g
Preparation of Sodium Acetate Solution:  Add components to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Sparge with 100% N 2 . Autoclave for 15 min at 15 psi pressure–
121°C.
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. Sparge with 100% N 2 . Autoclave for 15 min at 15 psi pressure–121°C.
Selenite/Tungstate Solution:
Composition per liter:
NaOH............................................................................................0.5g
Na 2 WO 4 ·2H 2 O...........................................................................4.0mg
Na 2 SeO 3 ·5H 2 O...........................................................................3.0mg
Preparation of Selenite/Tungstate Solution:  Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Sparge with 100% N 2 . Autoclave for 15 min at 15 psi pressure–121°C. Preparation of Medium: Add components, except sulfur, carbonate, sodium bicarbonate, and sodium acetate solution, to distilled/deionized water and bring volume to 990.0mL. Gently heat and bring to boiling. Boil for 1 min. Cool to room temperature under N 2  gas atmosphere. Add solid carbonate and bicarbonate. Ajust pH to 9.0–9.2. Dispense under N 2  gas atmosphere into culture vessels. Autoclave for 15 min at 15 psi pressure–121°C. Sterilize sulfur separately in screwcapped tubes by steaming in a water bath for 3 hr on each of 3 successive days. Aseptically add the sterilized sulfur to the medium. Mix thoroughly. Aseptically and anoxically add the sodium acetate solution. Mix thorougly.

Use: For the cultivation of Geoalkalibacter spp.