Desulfomonile Medium

[所属分类:培养基配方] [发布时间:2020-6-8] [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

Desulfomonile Medium

培养基配方
Composition per 1002.0mL:
Solution A...................................................................870.0mL
Solution C...................................................................100.0mL
Solution D....................................................................10.0mL
Solution E (Vitamin solution).................................................10.0mL
Solution F....................................................................10.0mL
Solution B (Trace elements solution SL-10) ....................................1.0mL
Solution G.....................................................................1.0mL
pH 6.8–7.0 at 25°C
Solution A:
Composition per 870.0mL:
Na2SO4 ........................................................................3.0g
NaCl...........................................................................1.0g
KCl............................................................................0.5g
MgCl2 ·6H2O ..................................................................0.4g
NH4Cl .........................................................................0.3g
KH2PO4 ........................................................................0.2g
CaCl2 ·2H2O .................................................................0.15g
Resazurin ....................................................................1.0mg
Preparation of Solution A: Add components to distilled/deionized
water and bring volume to 870.0mL. Mix thoroughly. Gently heat and
bring to boiling. Continue boiling for 3–4 min. Allow to cool to room
temperature while gassing under 80% N2 + 20% CO2 . Continue gas-
sing until pH reaches below 6.0. Seal the flask under 80% N2 + 20%
CO2 . Autoclave for 15 min at 15 psi pressure–121°C.
Solution B (Trace Elements Solution SL-10):
Composition per liter:
FeCl2 ·4H2O..................................................................1.5g
CoCl2 ·6H2O...............................................................190.0mg
MnCl2 ·4H2O ..............................................................100.0mg
ZnCl2 .....................................................................70.0mg
Na2MoO4 ·2H2O.............................................................36.0mg
NiCl2 ·6H2O...............................................................24.0mg
H3BO3 ......................................................................6.0mg
CuCl2 ·2H2O................................................................2.0mg
HCl (25% solution).........................................................10.0mL
Preparation of Solution B (Trace Elements Solution SL-10):
Add FeCl2 ·4H2O to 10.0mL of HCl solution. Mix thoroughly. Add dis-
tilled/deionized water and bring volume to 1.0L. Add remaining com-
ponents. Mix thoroughly. Gas under 100% N2 . Autoclave for 15 min at
15 psi pressure–121°C.
Solution C:
Composition per 100.0mL:
NaHCO3 ......................................................................2.5g
Preparation of Solution C:  Add NaHCO3 to distilled/deionized
water and bring volume to 100.0mL. Mix thoroughly. Filter sterilize.
Gas under 80% N2 + 20% CO2 .
Solution D:
Composition per 10.0mL:
Sodium pyruvate..............................................................4.0g
Preparation of Solution D:  Add sodium pyruvate to distilled/de-
ionized water and bring volume to 10.0mL. Mix thoroughly. Gas under
100% N2 . Autoclave for 15 min at 15 psi pressure–121°C.
Solution E (Vitamin Solution):
Composition per liter:
Nicotinamide...............................................................50.0mg
1,4-Naphthoquinone ........................................................20.0mg
Pyridoxine·HCl............................................................5.0mg
Calcium  DL -pantothenate..................................................5.0mg
Thioctic acid .............................................................5.0mg
p-Aminobenzoic acid........................................................5.0mg
Riboflavin ................................................................5.0mg
Thiamine·HCl .............................................................5.0mg
Biotin ....................................................................5.0mg
Folic acid.................................................................5.0mg
Vitamin B 12 ..............................................................5.0mg
Hemin......................................................................5.0mg
Preparation of Solution E (Vitamin Solution): Add 1,4-naphtho-
quinone and hemin to 10.0mL of 0.1N NaOH. Mix thoroughly. Add re-
maining components and bring volume to 1.0L with distilled/deionized
water. Mix thoroughly. Gas under 100% N2 . Autoclave for 15 min at
15 psi pressure–121°C.
Solution F:
Composition per 10.0mL:
Na2S·9H2O................................................................0.4g
Preparation of Solution F:  Add Na2S·9H2O to distilled/deionized
water and bring volume to 10.0mL. Mix thoroughly. Gas under 100%
N2 . Autoclave for 15 min at 15 psi pressure–121°C.
Solution G:
Composition per 10.0mL:
NaOH......................................................................0.5g
Na2WO4 ·2H2O............................................................4.0mg
Na2SeO3 ·5H2O...........................................................3.0mg
Preparation of Solution G:  Add components to distilled/deionized
water and bring volume to 10.0mL. Mix thoroughly. Gas under 100%
N2 . Autoclave for 15 min at 15 psi pressure–121°C.
Preparation of Medium: Aseptically and anaerobically combine
870.0mL of sterile solution A with 1.0mL of sterile solution B,
100.0mL of sterile solution C, 10.0mL of sterile solution D, 10.0mL of
sterile solution E, and 10.0mL of sterile solution F, in that order. Mix
thoroughly. Anaerobically distribute into sterile tubes or flasks under
100% N2 . Add 50.0mg/L of sodium dithionite prior to inoculation.
Use:  For the cultivation and maintenance of Desulfomonile tiedjei.