Carboxydothermus Medium

[所属分类:培养基配方] [发布时间:2020-4-7] [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

Carboxydothermus Medium

培养基配方
Composition per 1030.0mL:
MgCl2 ·6H2O.................................................0.52g
KCl..........................................................0.33g
KH2PO4 ......................................................0.33g
NH4Cl .......................................................0.33g
CaCl2 ·2H2O.................................................0.29g
Resazurin ...................................................0.5mg
Trace elements solution SL-4 ...............................10.0mL
Vitamin solution............................................10.0mL
Yeast extract solution......................................10.0mL
Na2S·9H2O solution.........................................10.0mL
pH 6.9 ± 0.2 at 25°C
Trace Elements Solution SL-4:
Composition per liter:
EDTA..........................................................0.5g
FeSO4 ·7H2O..................................................0.2g
Trace elements solution SL-6 ..............................100.0mL
Preparation of Trace Elements Solution SL-4:  Add components
to distilled/deionized water and bring volume to 1.0L. Mix thoroughly.
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.
Vitamin Solution:
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 Vitamin Solution:  Add components to distilled/
deionized water and bring volume to 1.0L. Mix thoroughly. Filter ster-
ilize. Sparge with 100% N2 .
Yeast Extract Solution:
Composition per 10.0mL:
Yeast extract...............................................10.0mg
Preparation of Yeast Extract Solution:  Add yeast extract to dis-
tilled/deionized water and bring volume to 10.0mL. Mix thoroughly.
Autoclave under 100% N2  for 15 min at 15 psi pressure–121°C.
Na2S·9H2O Solution:
Composition per 10.0mL:
Na2S·9H2O...........................................................0.7g
Preparation of Na2S·9H2O Solution: Prepare and dispense solu-
tion anaerobically under 100% N2 . Add Na2S·9H2O to distilled/deion-
ized water and bring volume to 10.0mL. Mix thoroughly. Adjust pH to
7.0. Autoclave for 15 min at 15 psi–121°C.
Preparation of Medium: Add components, except vitamin solu-
tion, yeast extract solution, and Na2S·9H2O solution, to distilled/deion-
ized water and bring volume to 1.0L. Mix thoroughly. Sparge under
100% N2  for 10 min. Autoclave under 100% N2  for 15 min at 15 psi
pressure–121°C. Aseptically and anaerobically combine with 10.0mL
of sterile vitamin solution, 10.0mL of sterile yeast extract solution, and
10.0mL of sterile Na2S·9H2O solution. Mix thoroughly. Pressurize in-
oculation flask with CO (carbon monoxide) gas at 2 bar pressure.
Caution:  Carbon monoxide is a toxic gas.
Use:  For the cultivationof Carboxydothermus hydrogenoformans.