Desulfovibrio sax Medium (DSMZ Medium 383a)

[所属分类:培养基配方] [发布时间:2020-6-10] [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

Desulfovibrio sax Medium
(DSMZ Medium 383a)

培养基配方
Composition per 1022.6mL:
Soultion A..............................................................930.0mL
Solution C...............................................................50.0mL
Solution E ..............................................................20.0mL
Solution D...............................................................10.0mL
Solution G...............................................................10.0mL
Solution B................................................................1.0mL
Solution F................................................................1.0mL
Vitamin B 12  solution ...................................................0.5mL
Yeast extract solution....................................................0.1mL
pH 7.3 at 25°C
Solution A:
Composition per 930.0mL:
NaCl.....................................................................21.0g
Na2SO4 ...................................................................3.0g
MgCl2 ·6H2O .............................................................3.0g
KCl.......................................................................0.5g
NH4Cl ....................................................................0.3g
KH2PO4 ...................................................................0.2g
CaCl2 ·2H2O ............................................................0.15g
Resazurin ...............................................................1.0mg
Preparation of Solution A : Add components to distilled/deionized
water and bring volume to 930.0mL. Mix thoroughly. Sparge with 80%
N2  + 20% CO2 gas until saturated. Autoclave for 15 min at 15 psi pres-
sure–121°C. Cool to 25°C.
Solution B:
Composition per liter:
FeCl2 ·4H2O...............................................................1.5g
H3BO3 ..................................................................300.0mg
CoCl2 ·6H2O............................................................190.0mg
MnCl2 ·4H2O ...........................................................100.0mg
ZnCl2 ...................................................................70.0mg
Na2MoO4 ·2H2O...........................................................36.0mg
NiCl2 ·6H2O.............................................................24.0mg
CuCl2 ·2H2O..............................................................2.0mg
HCl (25% solution)........................................................7.7mL
Preparation of Solution B: Add FeCl2 ·4H2O to 10.0mL of HCl so-
lution. Mix thoroughly. Add distilled/deionized water and bring vol-
ume to 1.0L. Add remaining components. Mix thoroughly. Sparge with
100% N2 . Autoclave for 15 min at 15 psi pressure–121°C.
Solution C:
Composition per 100.0mL:
NaHCO3 ...................................................................5.0g
Preparation of Solution C : Add NaHCO3 to distilled/deionized
water and bring volume to 100.0mL. Mix thoroughly. Sparge with
100% CO2 until saturated, approximately 20 min. Filter sterilize under
100% CO2 into a sterile, gas-tight 100.0mL screw-capped bottle.
Solution D:
Composition per 10.0mL:
Na-benzoate.............................................................0.5g
Preparation of Solution D:  Add Na-benzoate to distilled/deionized
water and bring volume to 10.0mL. Sparge with N2 . Filter sterilize. Store
anaerobically.
Solution E:
Composition per liter:
Pyridoxine-HCl.......................................................10.0mg
Thiamine-HCl·2H2O....................................................5.0mg
Riboflavin ...........................................................5.0mg
Nicotinic acid........................................................5.0mg
D -Ca-pantothenate....................................................5.0mg
p-Aminobenzoic acid...................................................5.0mg
Lipoic acid ..........................................................5.0mg
Biotin ...............................................................2.0mg
Folic acid............................................................2.0mg
Vitamin B 12 .........................................................0.1mg
Preparation of Solution E:  Add components to distilled/deionized
water and bring volume to 1.0L. Mix thoroughly. Sparge with 100%
N2 . Filter sterilize.
Solution F:
Composition per liter:
NaOH....................................................................0.5g
Na2SeO3 ·5H2O.........................................................3.0mg
Preparation of Solution F: Add components to distilled/deionized
water and bring volume to 1.0L. Mix thoroughly. Sparge with 100%
N2 . Filter sterilize.
Solution G:
Composition per 10.0mL:
Na2S·9H2O..............................................................0.4g
Preparation of Solution G:  Add Na2S·9H2O to distilled/deionized
water and bring volume to 10.0mL. Sparge with N2 . Autoclave for 15
min at 15 psi pressure–121°C. Cool to 25°C. Store anaerobically.
Vitamin B 12 Solution:
Composition per 100.0mL:
Vitamin B 12 .........................................................10.0mg
Vitamin B 12 Solution:  Add vitamin B 12 to distilled/deionized water
and bring volume to 100.0mL. Mix thoroughly. Sparge under 100% N2
gas for 3 min. Filter sterilize.
Yeast Extract Solution:
Composition per 10.0mL:
Yeast extract...........................................................1.0g
Preparation of Yeast Extract Solution: Add yeast extract to dis-
tilled/deionized water and bring volume to 10.0mL. Mix thoroughly.
Sparge with 100% N2 . Autoclave under 100% N2 for 15 min at 15 psi
pressure–121°C. Cool to room temperature.
Preparation of Medium:  Add solution B, solution C, solution D,
solution E, Vitamin B 12  solution, yeast extract solution, solution F, and
solution G to solution A in that order under N2 gas. Adjust the pH to
7.3.
Use: For the cultivation of Desulfotignum balticum (Desulfoarculus sp.).