Desulfotomaculum thermosapovorans 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 thermosapovorans Medium

培养基配方
Composition per 1015.0mL:
Solution A...............................................................900.0mL
Solution B...............................................................100.0mL
Solution C................................................................10.0mL
Solution D.................................................................5.0mL
Solution E ................................................................0.5mL
pH 7.0–7.2 at 25°C
Solution A:
Composition per 900.0mL:
NaCl.......................................................................8.0g
Na2SO4 ....................................................................3.0g
Sodium butyrate............................................................2.2g
NH4Cl .....................................................................1.0g
KCl........................................................................0.5g
Sodium acetate.............................................................0.5g
MgCl2 ·6H2O ..............................................................0.4g
Yeast extract..............................................................0.4g
KH2PO4 ....................................................................0.2g
CaCl2 ·2H2O .............................................................0.15g
Na2SeO3 ·5H2O..........................................................0.003mg
Trace elements solution SL-10.............................................1.5mL
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 compo-
nents. Mix thoroughly. Sparge with 100% N2 . Autoclave for 15 min at
15 psi pressure–121°C.
Preparation of Solution A: Add components to distilled/deionized
water and bring volume to 900.0mL. Mix thoroughly. Sparge with 80%
N2 + 20% CO2 .
Solution B:
Composition per 100.0mL:
NaHCO3 ..................................................................5.0g
Preparation of Solution B:  Add NaHCO3 to distilled/deionized
water and bring volume to 10.0mL. Mix thoroughly. Sparge with 100%
N2 . Autoclave for 15 min at 15 psi pressure–121°C.
Solution C:
Composition per liter:
Pyridoxine·HCl.........................................................10.0mg
p-Aminobenzoic acid.....................................................5.0mg
Lipoic acid ............................................................5.0mg
Nicotinic acid..........................................................5.0mg
Riboflavin .............................................................5.0mg
Thiamine·HCl ..........................................................5.0mg
Calcium  DL -pantothenate...............................................5.0mg
Biotin .................................................................2.0mg
Folic acid..............................................................2.0mg
Vitamin B 12 ...........................................................0.1mg
Preparation of Solution C:  Add components to distilled/deionized
water and bring volume to 1.0L. Mix thoroughly. Filter sterilize.
Solution D:
Composition per 5.0mL:
Na2S·9H2O...............................................................0.2g
Preparation of Solution D:  Add Na2S·9H2O to distilled/deionized
water and bring volume to 5.0mL. Mix thoroughly. Sparge with 100%
N2 . Autoclave for 15 min at 15 psi pressure–121°C. Before use, neu-
tralize to pH 7.0 with sterile HCl.
Solution E:
Composition per 1.0mL:
Na2S2O4 (sodium dithionite)..............................................20.0mg
Preparation of Solution E:  Add Na2S2O4  to distilled/deionized
water and bring volume to 1.0mL. Mix thoroughly. Sparge with 100%
N2 . Autoclave for 15 min at 15 psi pressure–121°C.
Preparation of Medium:  Prepare medium anaerobically under
80% N2 + 20% CO2 . Aseptically and anaerobically combine 900.0mL
of sterile solution A, 100.0mL of sterile solution B, 10.0mL of sterile
solution C, 5.0mL of sterile solution D, and 0.5 mL of sterile solution
E. Mix thoroughly. Adjust pH to 7.0–7.2.
Use: For the cultivation of Desulfotomaculum thermosapovorans.