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Haloanaerobium lacusroseus Medium (DSMZ Medium 764)
(DSMZ Medium 764)
培养基配方
Composition per liter:NaCl..........................................................................................200.0g
KCl............................................................................................... 4.0g
MgCl 2 ·6H 2 O..................................................................................2.0g
Yeast extract..................................................................................1.0g
NH 4 Cl ...........................................................................................1.0g
Na-acetate .....................................................................................1.0g
Trypticase™..................................................................................0.5g
K 2 HPO 4 .........................................................................................0.3g
KH 2 PO 4 .........................................................................................0.3g
CaCl 2 ·2H 2 O...................................................................................0.2g
Resazurin ..................................................................................0.001g
Glucose solution ....................................................................100.0mL
NaHCO 3 solution .....................................................................50.0mL
Na 2 S·9H 2 O solution.................................................................10.0mL
Dithionite solution .....................................................................5.0mL
Trace elements soslution SL-6...................................................1.0mL
pH 7.0 ± 0.2 at 25°C
Glucose Solution:
Composition per 100.0mL:
Glucose .......................................................................................17.4g
Preparation of Glucose Solution: Add glucose to distilled/deionized water and bring volume to 100.0mL. Mix thoroughly. Filter sterilize.
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. 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....................................................................................0.2g
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.
Neutralize to pH 7.0 with sterile HCl.
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.
Dithionite Solution
Composition per 10.0mL:
Na-dithionite..............................................................................2.0mg
Preparation of Dithionite Solution: Add Na-dithionite to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Sparge with 100% N 2 . Filter sterilize.
Preparation of Medium: Prepare and dispense medium under 80% N 2 + 20% CO 2 gas atmosphere. Add components, except glucose solution, NaHCO 3 solution, dithionite solution, and Na 2 S·9H 2 O solution, to distilled/deionized water and bring volume to 835.0mL. Mix thoroughly. Adjust pH to 7.0. Gently heat and bring to boiling. Cool while sparging with 80% N 2 + 20% CO 2 . Distribute into Hungate tubes under 80% N 2 + 20% CO 2 . Autoclave for 15 min at 15 psi pressure–121°C. Cool to room temperature. Aseptically and anaerobically inject glucose solution (0.25mL per 10mL medium), dithionite solution (0.05mL per 10mL medium), NaHCO 3 solution (0.5mL per 10mL medium), and Na 2 S·9H 2 O solution (0.1mL per 10mL medium). Aseptically and anaerobically distribute into sterile tubes or bottles.
Use: For the cultivation of Halanaerobium lacusrosei (Haloanaerobium lacusroseus).