DMA Medium

[所属分类:培养基配方] [发布时间:2020-6-22] [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

DMA Medium

培养基配方
Composition per liter:
NaHCO3 .....................................................................3.0g
KH2PO4 ....................................................................0.85g
K2HPO4 .....................................................................0.8g
NH4Cl ......................................................................0.5g
FeSO4 ·7H2O...............................................................1.0mg
Resazurin .................................................................0.5mg
Glucose solution.........................................................100.0mL
Wolfe’s mineral solution.................................................10.0mL
Wolfe’s vitamin solution.................................................10.0mL
Na2S·9H2O solution.......................................................10.0mL
MgSO4 ·7H2O solution.....................................................10.0mL
Glucose Solution:
Composition per 100.0mL:
D -Glucose.................................................................2.0g
Preparation of Glucose Solution:  Add glucose to distilled/deion-
ized water and bring volume to 100.0mL. Mix thoroughly. Sparge with
100% N2 . Autoclave for 15 min at 15 psi pressure–121°C.
Wolfe’s Mineral Solution:
Composition per liter:
MgSO4 ·7H2O................................................................3.0g
Nitrilotriacetic acid.......................................................1.5g
NaCl........................................................................1.0g
MnSO4 ·2H2O................................................................0.5g
CoCl2 ·6H2O................................................................0.1g
ZnSO4 ·7H2O................................................................0.1g
CaCl2 ·2H2O ...............................................................0.1g
FeSO4 ·7H2O................................................................0.1g
NiCl2 ·6H2O..............................................................0.025g
KAl(SO4)2 ·12H2O..........................................................0.02g
CuSO4 ·5H2O...............................................................0.01g
H3BO3 .....................................................................0.01g
Na2MoO4 ·2H2O.............................................................0.01g
Na2SeO3 ·5H2O.............................................................0.3mg
Preparation of Wolfe’s Mineral Solution: Add nitrilotriacetic
acid to 500.0mL of distilled/deionized water. Adjust pH to 6.5 with
KOH. Add remaining components. Add distilled/deionized water to
1.0L. Sparge with 100% N2 . Adjust pH to 6.8. Filter sterilize.
Wolfe’s Vitamin Solution:
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 Wolfe’s Vitamin Solution:  Add components to
distilled/deionized water and bring volume to 1.0L. Mix thoroughly.
Filter sterilize.
MgSO4 ·7H2O Solution:
Composition per 10.0mL:
MgSO4 ·7H2O...............................................................0.25g
Preparation of MgSO4 ·7H2O Solution:  Add MgSO4 ·7H2O to
distilled/deionized water and bring volume to 10.0mL. Mix thorough-
ly. Sparge with 100% N2 . Autoclave for 15 min at 15 psi pressure–
121°C.
Na2S·9H2O Solution:
Composition per 10.0mL:
Na2S·9H2O.................................................................0.3g
Preparation of Na2S·9H2O Solution:  Add Na2S·9H2O to dis-
tilled/deionized water and bring volume to 10.0mL. Mix thoroughly.
Sparge with 100% N2 . Autoclave for 15 min at 15 psi pressure–121°C.
Before use, neutralize to pH 7.0 with sterile HCl.
Preparation of Medium: Prepare and dispense medium under 80%
N2 + 20% CO2 gas mixture. Add components, except glucose solution,
Wolfe’s mineral solution, Wolfe’s vitamin solution, Na 2 S·9H 2 O solu-
tion, and MgSO4 ·7H2O solution, to distilled/deionized water and bring
volume to 860.0mL. Mix thoroughly. Sparge with 80% N2 + 20% CO2
gas mixture for 30 min. Autoclave for 15 min at 15 psi pressure–121°C.
Aseptically and anaerobically add 100.0mL of sterile glucose solution,
10.0mL of sterile Wolfe’s mineral solution, 10.0mL of sterile Wolfe’s
vitamin solution, 10.0mL of sterile Na2S·9H2O solution, and 10.0mL
of sterile MgSO4 ·7H2O solution. Mix thoroughly. Aseptically and an-
aerobically distribute into sterile tubes or bottles.
Use: For the cultivation of unclassified bacterium DSMZ 8827.