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 1051.0mL:
Elemental sulfur slurry ...................................................10.0g
Solution 1..................................................................1.0L
Solution 3................................................................40.0mL
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:
NaCl......................................................................20.0g
MgCl2 ·6H2O ..............................................................3.0g
KH2PO4 ....................................................................1.0g
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
pH 6.5 ± 0.2 at 25°C
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 marine Desulfuromonas spe-
cies.