Desulfuromonas Medium

[所属分类:培养基配方] [发布时间:2020-6-11] [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

Desulfuromonas Medium

培养基配方
Composition per 1031.0mL:
Elemental sulfur slurry..................................................10.0g
Solution 1................................................................1.0L
Solution 3..............................................................20.0mL
Solution 4...............................................................6.0mL
Solution 5...............................................................5.0mL
Solution 2...............................................................1.0mL
pH 7.2 ± 0.2 at 25°C
Solution 1:
Composition per liter:
KH2PO4 ...................................................................1.0g
MgCl2 ·6H2O..............................................................0.4g
NH4Cl ....................................................................0.3g
CaCl2 ·2H2O .............................................................0.1g
HCl (2N solution) .......................................................4.0mL
Preparation of Solution 1: Add components to distilled/deionized
water and bring volume to 1.0L. Mix thoroughly. Autoclave for 15 min
at 15 psi pressure–121°C. Cool to 25°C.
Solution 2:
Composition per liter:
Disodium EDTA.............................................................5.2g
CoCl2 ·6H2O..............................................................1.9g
FeCl2 ·4H2O..............................................................1.5g
MnCl2 ·4H2O .............................................................1.0g
ZnCl2 ....................................................................0.7g
H3BO3 ...................................................................0.62g
Na2MoO4 ·2H2O...........................................................0.36g
NiCl2 ·6H2O.............................................................0.24g
CuCl2 ·2H2O.............................................................0.17g
Preparation of Solution 2: Add components to distilled/deionized
water and bring volume to 1.0L. Mix thoroughly. Adjust pH to 6.5. Au-
toclave for 15 min at 15 psi pressure–121°C. Cool to 25°C.
Solution 3:
Composition per 100.0mL:
NaHCO3 ..................................................................10.0g
Preparation of Solution 3: Add NaHCO3 to distilled/deionized
water and bring volume to 100.0mL. Mix thoroughly. Autoclave for 15
min at 15 psi pressure–121°C. Cool to 25°C.
Solution 4:
Composition per 100.0mL:
Na2S·9H2O...............................................................5.0g
Preparation of Solution 4: Add Na2S·9H2O to distilled/deionized
water and bring volume to 100.0mL. Mix thoroughly. Autoclave for 15
min at 15 psi pressure–121°C. Cool to 25°C.
Solution 5:
Composition per 200.0mL:
Pyridoxamine·HCl......................................................0.01g
Nicotinic acid.........................................................4.0mg
p-Aminobenzoic acid....................................................2.0mg
Thiamine...............................................................2.0mg
Cyanocobalamin.........................................................1.0mg
Pantothenic acid.......................................................1.0mg
Biotin.................................................................0.5mg
Preparation of Solution 5: Add components to distilled/deionized
water and bring volume to 200.0mL. Mix thoroughly. Filter sterilize.
Elemental Sulfur Slurry:
Composition per 10.0g:
Sulfur flowers..........................................................10.0g
Preparation of Elemental Sulfur Slurry: Add highly purified
sulfur flowers to a mortar and grind to a fine powder. Add sufficient
distilled/deionized water to produce a slurry. Distribute into 100.0mL
screw-capped bottles in 20.0mL volumes. Autoclave for 30 min at 10
psi pressure–115°C. Decant supernatant solution. Reserve sulfur slur-
ry.
Preparation of Medium: To 1.0L of cooled, sterile solution 1,
aseptically add 1.0mL of sterile solution 2, 40.0mL of sterile solution
3, 6.0mL of sterile solution 4, and 5.0mL of sterile solution 5. Mix thor-
oughly. Adjust pH to 7.2. Aseptically distribute into sterile 50.0mL
screw-capped bottles. Fill bottles completely with medium except for
a pea-sized air bubble. Aseptically add a pea-sized piece of sulfur slur-
ry to each 50.0mL of medium.
Use: For the isolation and cultivation of freshwater Desulfuromonas
species.