Desulfotomaculum sapomandens Medium

[所属分类:培养基配方] [发布时间:2020-6-9] [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

Desulfotomaculum sapomandens Medium

培养基配方
Composition per 1019.7mL:
Solution A.................................................................966.0mL
Solution B..................................................................20.0mL
Solution C..................................................................10.0mL
Solution D..................................................................10.0mL
Solution F..................................................................10.0mL
Solution H...................................................................1.7mL
Solution G...................................................................1.0mL
Solution E ..................................................................1.0mL
pH 7.2–7.5 at 25°C
Solution A:
Composition per 966.0mL:
Na2SO4 .......................................................................3.0g
NaCl..........................................................................1.0g
MgCl2 ·6H2O .................................................................0.4g
NH4Cl ........................................................................0.3g
KH2PO4 .......................................................................0.2g
CaCl2 ·2H2O ................................................................0.15g
Resazurin....................................................................1.0mg
Trace elements solution SL-10................................................1.0mL
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 Trace Elements Solution SL-10:  Add FeCl2 ·4H2O
to 10.0mL of HCl solution. Mix thoroughly. Add distilled/deionized
water and bring volume to 1.0L. Add remaining components. Mix thor-
oughly. Gas under 100% N2 . Autoclave for 15 min at 15 psi pressure–
121°C.
Solution B:
Composition per 20.0mL:
NaHCO3 ......................................................................1.0g
Preparation of Solution B: Add NaHCO3 to distilled/deionized
water and bring volume to 20.0mL. Mix thoroughly. Gas under 100%
N2 . Autoclave for 15 min at 15 psi pressure–121°C.
Solution C:
Composition per 10.0mL:
Ethanol.....................................................................1.0mL
Preparation of Solution C: Add ethanol to distilled/deionized wa-
ter and bring volume to 10.0mL. Mix thoroughly. Filter sterilize.
Solution D:
Composition per 10.0mL:
Sodium benzoate..............................................................0.7g
Preparation of Solution D:  Add sodium benzoate to distilled/de-
ionized water and bring volume to 10.0mL. Mix thoroughly. Filter ster-
ilize.
Solution E:
Composition per 1.0mL:
Rumen fluid, clarified.....................................................1.0mL
Preparation of Solution E:  Gas under 100% N2 . Autoclave for 15
min at 15 psi pressure–121°C.
Solution F:
Composition per liter:
Pyridoxine·HCl...........................................................10.0mg
Calcium  DL -pantothenate..................................................5.0mg
Lipoic acid ...............................................................5.0mg
Nicotinic acid.............................................................5.0mg
p-Aminobenzoic acid........................................................5.0mg
Riboflavin ................................................................5.0mg
Thiamine·HCl .............................................................5.0mg
Biotin ....................................................................2.0mg
Folic acid.................................................................2.0mg
Vitamin B 12 ..............................................................0.1mg
Preparation of Solution F: Add components to distilled/deionized
water and bring volume to 1.0L. Mix thoroughly. Gas under 100% N 2 .
Filter sterilize.
Solution G:
Composition per 1.0mL:
Sodium dithionite.........................................................0.025g
Preparation of Solution G: Add sodium dithionite to distilled/de-
ionized water and bring volume to 1.0mL. Mix thoroughly. Gas under
100% N2 . Filter sterilize.
Solution H:
Composition per 10.0mL:
Na2S·9H2O.................................................................0.3g
Preparation of Solution H:  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.
Preparation of Medium: Aseptically and anaerobically combine in
the following order: 966.0mL of sterile solution A with 20.0.0mL of
sterile solution B, 10.0mL of sterile solution C, 10.0mL of sterile solu-
tion D, 1.0mL of sterile solution E, 10.0mL of sterile solution F, 1.0mL
of sterile solution G, and 1.7mL of sterile solution H. Mix thoroughly.
Use:  For the cultivation and maintenance of Desulfotomaculum sapo-
mandens.