Chloroflexus aggregans Medium (DSMZ Medium 87a)

[所属分类:培养基配方] [发布时间:2020-4-20] [发布人:] [阅读次数:] [返回]

山东拓普生物工程有限公司

Shandong Tuopu Biol-Engineering Co.,Ltd

Chloroflexus aggregans Medium

培养基配方
(DSMZ Medium 87a)
Composition per 1061.0mL:
Yeast extract..................................................1.0g
Glycyl-glycine ................................................1.0g
NaNO3 .........................................................0.5g
Na2HPO4 ·7H2O.................................................0.1g
MgSO4 ·7H2O...................................................0.1g
KNO3 ..........................................................0.1g
NaCl...........................................................0.1g
CaCl2 ·2H2O..................................................0.05g
Neutralized sulfide solution.................................11.0mL
Ferric citrate solution.......................................5.0mL
Trace elements solution SL-6 .................................1.0mL
pH 8.2 ± 0.2 at 25°C
Ferric Citrate Solution:
Composition per 100.0mL:
Ferric citrate................................................0.1mg
Preparation of Ferric Citrate Solution : Add ferric citrate to dis-
tilled/deionized water and bring volume to 100.0mL. Mix thoroughly.
Sparge under 100% N2 gas for 3 min.
Trace Elements Solution SL-6:
Composition per liter:
MnCl2 ·4H2O ...................................................0.5g
H3BO3 ..........................................................0.3g
CoCl2 ·6H2O....................................................0.2g
ZnSO4 ·7H2O....................................................0.1g
Na2MoO4 ·2H2O.................................................0.03g
NiCl2 ·6H2O...................................................0.02g
CuCl2 ·2H2O...................................................0.01g
Preparation of Trace Elements Solution SL-6: Add components
to distilled/deionized water and bring volume to 1.0L. Mix thoroughly.
Neutralized Sulfide Solution:
Composition per 100.0mL:
Na2S·9H2O.......................................................1.5g
Preparation of Neutralized Sulfide Solution : Add Na2S·9H20
to distilled/deionized water in a 250mL screw-capped bottle fitted with
a butyl rubber septum and bring volume to 100.0mL. Add a magnetic
stir bar. Mix thoroughly. Sparge under 100% N2 gas for 3 min. Auto-
clave for 15 min at 15 psi pressure–121°C. Cool to room temperature.
Adjust pH to about 7.3 with sterile 2M H2SO4 . Do not open the bottle
to add H2SO4 ; use a sterile syringe. Stir the solution continuously to
avoid precipitation of elemental sulfur. The final solution should be
clear and yellow in color.
Preparation of Medium: Add components, except neutralized sul-
fide solution, to distilled/deionized water and bring volume to
1050.0mL. Mix thoroughly. Adjust pH to 8.2. Gently heat and bring to
boiling. Continue boiling for 3–4 min under 100% N2 . Distribute
90.0mL of medium into 100mL screw-capped bottles with rubber septa
under 100% N2 . Autoclave for 15 min at 15 psi pressure–121°C. Cool
to room temperature. Using a sterile syringe, inject 1.0mL of neutral-
ized sulfide solution into each bottle. Incubate the culture at 50°C at a
light intensity of 300–500 lux. For heavy cell suspension supplement
periodically with sterile yeast extract solution to yield a final concen-
tration of 0.1%.
Use: For the growth and maintenance of Chloroflexus aggregans and
Roseiflexus castenholzii.