HESP1/SR1/TMC4/LUP Medium (DSMZ Medium 860)

[所属分类:培养基配方] [发布时间:2021-4-6] [发布人: ] [阅读次数:] [返回]

HESP1/SR1/TMC4/LUP Medium

(DSMZ Medium 860)

培养基配方

Composition per 1090.0mL:
Yeast extract..................................................................................1.0g
NH 4 Cl ...........................................................................................1.0g
NaCl..............................................................................................0.6g
Cysteine-HCl·H 2 O........................................................................0.5g
K 2 HPO 4 .........................................................................................0.3g
KH 2 PO 4 .........................................................................................0.3g
MgCl 2 ·6H 2 O .................................................................................0.2g
CaCl 2 ·2H 2 O ..................................................................................0.1g
KCl................................................................................................0.1g
Resazurin...................................................................................0.5mg
NaHCO 3  solution.....................................................................60.0mL
Substrate solution.....................................................................20.0mL
Na 2 S·9H 2 O solution.................................................................10.0mL
Trace elements solution SL-10..................................................1.5mL
pH 7.0 ± 0.2 at 25°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. Mix thoroughly. Sparge with 100% N 2 . Autoclave for 15 min at 15 psi pressure–121°C.
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 2 S·9H 2 O Solution:
Composition per 10.0mL:
Na 2 S·9H 2 O....................................................................................0.3g
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. Sparge with N 2 . Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C. Store anaerobically.
Substrate Solution:
Composition per 10.0mL:
Casamino acids.............................................................................0.5g
Preparation of Substrate Solution:  Add casamino acids to distilled/deionized water and bring volume to 10.0mL. Sparge with N 2 . Filter sterilize.
Preparation of Medium: Prepare and dispense medium under 80% N 2  + 20% CO 2 . Add components, except NaHCO 3 solution, Na 2 S·9H 2 O solution, and substrate solution, to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Adjust pH to 7.0. Gently heat and bring to boiling. Boil for 10 min. Cool to room temperature while sparging with 80% N 2  + 20% CO 2 . Adjust pH to 6.0. Dispense under 80% N 2  + 20% CO 2  into tubes or bottles. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C. Aseptically and anaerobically add 0.6mL sterile NaHCO 3  solution per 10.0mL medium, 0.1mL Na 2 S·9H 2 O solution per 10.0mL medium, and 0.2mL substrate solution per 10.0mL medium. Final pH is 7.0.
Use:  For the cultivation of Clostridium peptidivorans DSM 12505.