Desulfobacula toluolica Medium (DSMZ Medium 383b)

[所属分类:培养基配方] [发布时间:2020-6-5] [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

Desulfobacula toluolica Medium
(DSMZ Medium 383b)

培养基配方
Composition per 1013.5mL:
Soultion A.................................................................930.0mL
Solution C..................................................................50.0mL
Solution D..................................................................10.0mL
Solution E .................................................................10.0mL
Solution G..................................................................10.0mL
Solution B...................................................................1.0mL
Solution F...................................................................1.0mL
Sodium-tungstate solution....................................................1.0mL
Vitamin B 12  solution ......................................................0.5mL
pH 7.0 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:
Na-EDTA.......................................................................5.2g
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 7.7mL of HCl so-
lution. Mix thoroughly. Add distilled/deionized water and bring vol-
ume to 1.0L. Add remaining components. Mix thoroughly. Adjust pH
to 6.0. 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 minutes. Filter sterilize
under 100% CO2 into a sterile, gas-tight 100.0mL screw-capped bottle.
Solution D:
Composition per 10.0mL:
Na-benzoate.................................................................0.4g
Preparation of Solution D:  Add Na-benzoate to distilled/deion-
ized water and bring volume to 10.0mL. Sparge with N2 . Filter steril-
ize. 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% N 2
gas for 3 min. Filter sterilize.
Selenite-Tungstate Solution:
Composition per liter:
NaOH.....................................................................0.5g
Na2WO4 ·2H2O...........................................................4.0mg
Na2SeO3 ·5H2O..........................................................3.0mg
Preparation of Selenite-Tungstate Solution: Add components
to distilled/deionized water and bring volume to 1.0L. Mix thoroughly.
Sparge with 100% N2 . Filter sterilize.
Preparation of Medium:  Add solution B, solution C, solution D,
solution E, Vitamin B 12  solution, solution F, selenite-tungstate solution,
and solution G to solution A in that order under N2 gas. Adjust the pH
to 7.0.
Use: For the cultivation of Desulfobacula toluolica.