Haloanaerobacter chitinovorans Medium

[所属分类:培养基配方] [发布时间:2021-3-24] [发布人: ] [阅读次数:] [返回]

Haloanaerobacter chitinovorans Medium

培养基配方

Composition per 1001.0mL:
NaCl............................................................................................10.0g
MgSO 4 ·7H 2 O................................................................................9.6g
MgCl 2 ·6H 2 O..................................................................................7.0g
KCl................................................................................................3.8g
Na 2 CO 3 .........................................................................................1.0g

NH 4 Cl ...........................................................................................1.0g

Yeast extract..................................................................................1.0g
CaCl 2 ·2H 2 O ..................................................................................0.5g
K 2 HPO 4 ·3H 2 O .............................................................................0.4g
Resazurin..................................................................................0.001g
Na 2 CO 3  solution.......................................................................20.0mL
Substrate solution.....................................................................20.0mL
Na 2 S·9H 2 O solution.................................................................10.0mL
L -Cysteine·HCl·H 2 O solution..................................................10.0mL
Trace elements solution SL-6....................................................1.0mL
pH 7.2 ± 0.2 at 25°C
Na 2 CO 3 Solution:
Composition per 20.0mL:
Na 2 CO 3 .........................................................................................3.0g
Preparation of Na 2 CO 3 Solution:  Add Na 2 CO 3 to distilled/deionized water and bring volume to 20.0mL. Mix thoroughly. Filter sterilize. Sparge with 100% N 2 .
Substrate Solution:
Composition per 20.0mL:
Glucose or N-acetylglucosamine..................................................5.0g
Preparation of Substrate Solution:  Add glucose or N-acetylglucosamine to distilled/deionized water and bring volume to 20.0mL. Mix thoroughly. Sparge with 100% N 2 . Autoclave for 15 min at 15 psi pressure–121°C.
Na 2 S·9H 2 O Solution:
Composition per 10.0mL:
Na 2 S·9H 2 O....................................................................................0.5g
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. Sparge with 100% N 2 . Autoclave for 15 min at 15 psi pressure–121°C. Before use, neutralize to pH 7.0 with sterile HCl.
L -Cysteine·HCl·H 2 O Solution:
Composition per 10.0mL:
L -Cysteine·HCl·H 2 O.....................................................................0.5g
Preparation of  L -Cysteine·HCl·H 2 O 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-6:
Composition per liter:
MnCl 2 ·4H 2 O .................................................................................0.5g
H 3 BO 3 ...........................................................................................0.3g
CoCl 2 ·6H 2 O..................................................................................0.2g
ZnSO 4 ·7H 2 O................................................................................ 0.1g
Na 2 MoO 4 ·2H 2 O......................................................................... 0.03g
NiCl 2 ·6H 2 O.................................................................................0.02g
CuCl 2 ·2H 2 O................................................................................0.01g
Preparation of Trace Elements Solution SL-6: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly.
Preparation of Medium: Prepare and dispense medium under 100% N 2 . Add components, except Na 2 CO 3  solution, Na 2 S·9H 2 O solution, and  L -cysteine·HCl·H 2 O solution, to distilled/deionized water and bring volume to 960.0mL. Mix thoroughly. Adjust pH to 7.2. Gently heat and bring to boiling. Cool to room temperature while sparging with 100% N 2 . Autoclave for 15 min at 15 psi pressure–121°C. Aseptically and anaerobically add 20.0mL of sterile NaHCO 3  solution, 10.0mL of sterile Na 2 S·9H 2 O solution, and 10.0mL of sterile  L cysteine·HCl·H 2 O solution. Mix thoroughly. Aseptically distribute into sterile tubes or flasks.
Use: For the cultivation of Haloanaerobacter chitinovorans.