Gracilibacillus Medium (DSMZ Medium 802)

[所属分类:培养基配方] [发布时间:2021-3-19] [发布人: ] [阅读次数:] [返回]

Gracilibacillus Medium

(DSMZ Medium 802)

培养基配方

Composition per liter:
NaCl..........................................................................................100.0g
MgSO 4 ·7H 2 O..............................................................................20.0g
Tris-HCl......................................................................................12.0g
KCl................................................................................................2.0g
Yeast extract..................................................................................2.0g
Trypticase™ peptone....................................................................2.0g
NaBr..............................................................................................0.1g
Trace metals solution.................................................................2.0mL

Phosphate solution...................................................................10.0mL

Glucose solution ......................................................................10.0mL
Calcium chloride solution..........................................................5.0mL
Iron chloride manganese chloride solution................................2.0mL
pH 7.8 ± 0.2 at 25°C
Trace Metals Solution:
Composition per liter:
FeCl 2 ·4H 2 O...................................................................................2.0g
CoCl 2 ·6H 2 O ...........................................................................250.0mg
MnCl 2 ·4H 2 O...........................................................................100.0mg
ZnCl 2 ........................................................................................70.0mg
NiCl 2 ·6H 2 O..............................................................................70.0mg
AlCl 3 ·6H 2 O..............................................................................60.0mg
Na 2 MoO 4 ·2H 2 O .......................................................................40.0mg
H 3 BO 3 ........................................................................................6.0mg
Na 2 WO 4 ·2H 2 O...........................................................................6.0mg
CuCl 2 ·2H 2 O ...............................................................................2.0mg
HCl (32% solution)..................................................................10.0mL
Preparation of Trace Metals Solution: 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.
Phosphate Solution:
Composition per liter:
KH 2 PO 4 .......................................................................................50.0g
Preparation of Phosphate Solution:  Add KH 2 PO 4 to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C.
Calcium Chloride Solution:
Composition per 10.0mL:
CaCl 2 ·2H 2 O...................................................................................0.1g
Preparation of Calcium Chloride Solution:  Add CaCl 2 ·2H 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. Cool to 25°C.
Iron Chloride Manganese Chloride Solution:
Composition per 10.0mL:
FeCl 2 ·4H 2 O...................................................................................0.2g
MnCl 2 ·4H 2 O..................................................................................0.2g
Preparation of Iron Chloride Manganese Chloride 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. Cool to 25°C.
Glucose Solution:
Composition per 10.0mL:
Glucose .........................................................................................2.0g
Preparation of Glucose Solution:  Add glucose to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Sparge with 100% N 2 . Filter sterilize.
Preparation of Medium: Add components, except glucose solution, phosphate solution, calcium chloride solution, and iron chloride manganese chloride solution, to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Adjust pH to 7.8. Autoclave for 15 min at 15 psi pressure–121°C. Cool to room temperature. Aseptically add 10.0mL glucose solution, 10.0mL phosphate solution, 5.0mL calcium chloride solution, and 2.0mL iron chloride manganese chloride solution. Mix thoroughly. Aseptically and anaerobically distribute into sterile tubes or bottles.
Use:  For the cultivation of Gracilibacillus spp.