GOL CHI1 Medium (DSMZ Medium 298c)

[所属分类:培养基配方] [发布时间:2021-3-18] [发布人: ] [阅读次数:] [返回]

GOL CHI1 Medium

(DSMZ Medium 298c)

培养基配方

Composition per liter:
NaCl..............................................................................................1.0g
KCl................................................................................................0.5g
MgCl 2 ·6H 2 O .................................................................................0.4g
NH 4 Cl .........................................................................................0.25g
KH 2 PO 4 .........................................................................................0.2g
CaCl 2 ·2H 2 O ................................................................................0.15g
Resazurin ...................................................................................1.0mg

NaHCO 3  solution.....................................................................10.0mL

Butanediol solution..................................................................10.0mL
Na 2 S·9H 2 O solution.................................................................10.0mL
Vitamin solution.......................................................................10.0mL
Quinic acid solution.................................................................10.0mL
Trace elements solution SL-10 ..................................................1.0mL
pH 7.2 ± 0.2 at 25°C
Na 2 S·9H 2 O Solution :
Composition per 10.0mL:
Na 2 S·9H 2 O..................................................................................0.36g
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 ........................................................................................2.5g
Preparation of NaHCO 3 Solution:  Add NaHCO 3 to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Sparge with 80% N 2 + 20% CO 2 . Filter sterilize.
Butanediol Solution:
Composition per 10.0mL:
2,3 butanediol................................................................................0.9g
Preparation of Butanediol Solution: Add butanediol to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Sparge with 100% N 2 . Filter sterilize.
Trace Elements Solution SL-10:
Composition per liter:
FeCl 2 ·4H 2 O...................................................................................1.5g
H 3 BO 3 ....................................................................................300.0mg
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
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 80% N 2 + 20% CO 2 . Autoclave for 15 min at 15 psi pressure–121°C.
Vitamin Solution:
Composition per liter:
Pyridoxine-HCl........................................................................10.0mg
Thiamine-HCl·2H 2 O..................................................................5.0mg
Riboflavin ..................................................................................5.0mg
Nicotinic acid.............................................................................5.0mg
D -Ca-pantothenate......................................................................5.0mg
p-Aminobenzoic acid.................................................................5.0mg
Lipoic acid .................................................................................5.0mg
Biotin .........................................................................................2.0mg
Folic acid....................................................................................2.0mg
Vitamin B 12 ................................................................................0.1mg
Preparation of Vitamin Solution:  Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Sparge with 80% H 2 + 20% CO 2 . Filter sterilize.
Quinic Acid Solution:
Composition per 10.0mL:
Quinic acid....................................................................................1.0g
Preparation of Quinic Acid Solution:  Add quinic acid to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Adjust pH to 7.0. Sparge with 80% N 2 + 20% CO 2 . Filter sterilize.

Preparation of Medium: Prepare and dispense medium under 80% N 2 + 20% CO 2 gas atmosphere. Add components, except NaHCO 3  solution, butanediol solution, Na 2 S·9H 2 O solution, vitamin solution, quinic acid solution, and trace elements solution SL-10, to distilled/deionized water and bring volume to 949.0mL. Mix thoroughly. Adjust pH to 7.2. Sparge with 80% N 2 + 20% CO 2 . Autoclave for 15 min at 15 psi pressure–121°C. Aseptically and anaerobically add 10.0mL NaHCO 3  solution, 10.0mL butanediol solution, 10.0mL Na 2 S·9H 2 O solution, 10.0mL vitamin solution, 10.0mL quinic acid solution, and 1.0mL trace elements solution SL-10. Mix thoroughly. Aseptically and anaerobically distribute into sterile tubes or bottles. After inoculation, flush and repressurize the gas head space of culture bottles with sterile 80% N 2 + 20% CO 2 to 1 bar overpressure.
Use:  For the cultivation of Propionivibrio limicola.