FlGlyM Medium

[所属分类:培养基配方] [发布时间:2021-3-1] [发布人: ] [阅读次数:] [返回]

FlGlyM Medium
(DSMZ Medium 298b)

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.....................................................................50.0mL

Na 2 S·9H 2 O solution.................................................................10.0mL
Na-glyoxalate solution.............................................................10.0mL
L -Cysteine solution..................................................................10.0mL
Na-acetate solution ....................................................................7.0mL
Trace elements solution SL-10 ..................................................1.0mL
Vitamin solution.........................................................................0.5mL
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 100.0mL:
NaHCO 3 ......................................................................................10.0g
Preparation of NaHCO 3 Solution:  Add NaHCO 3 to distilled/deionized water and bring volume to 100.0mL. Mix thoroughly. Sparge with 80% N 2 + 20% CO 2 . Filter sterilize.
Na-Acetate Solution:
Composition per 10.0mL:
Na-acetate ...................................................................................0.25g
Preparation of Na-Acetate Solution:  Add Na-acetate to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Sparge with 100% N 2 . Filter sterilize.
Na-Glyoxalate Solution:
Composition per 10.0mL:
Na-glyoxalate................................................................................0.6g
Preparation of Na-Glyoxalate Solution:  Add Na-glyoxalate to
distilled/deionized water and bring volume to 10.0mL. Mix thorough-
ly. Sparge with 100% N 2 . Filter sterilize.
L -Cysteine Solution:
Composition per 10.0mL:
L -Cysteine·HCl·H 2 O .....................................................................0.3g
Preparation of  L -Cysteine Solution:  Add  L -cysteine·HCl·H 2 O 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
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 hydrochloride ......................................................300.0mg
Thiamine-HCl·2H 2 O..............................................................200.0mg
Nicotinic acid.........................................................................200.0mg
Vitamin B 12 ............................................................................100.0mg
Calcium pantothenate ............................................................100.0mg
p-Aminobenzoic acid...............................................................80.0mg
Folic Acid................................................................................30.0mg
D (+)-Biotin...............................................................................20.0mg
α-Lipoic acid...........................................................................10.0mg
Preparation of Vitamin Solution: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Filter sterilize.
Preparation of Medium: Prepare and dispense medium under 80% N 2 + 20% CO 2 gas atmosphere. Add components, except NaHCO 3  solution, Na 2 S·9H 2 O solution,  L -cysteine solution, Na-acetate solution, Naglyoxalate solution, vitamin solution, and trace elements solution SL- 10, to distilled/deionized water and bring volume to 979.0mL. 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 Na 2 S·9H 2 O solution, 10.0mL  L - cysteine solution, 7.0mL Na-acetate solution, 10.0mL Na-glyoxalate solution, 1.0mL trace elements solution SL-10, and 0.5mL vitamin solution. 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 Syntrophobotulus glycolicus.