Dethiosulfovibrio peptidovorans Medium (DSMZ Medium 786)

[所属分类:培养基配方] [发布时间:2020-6-12] [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

Dethiosulfovibrio peptidovorans Medium
(DSMZ Medium 786)

培养基配方
Composition per 1085.0mL:
NaCl......................................................................30.0g
Trypticase™................................................................5.0g
MgCl2 ·6H2O...............................................................3.0g
Yeast extract..............................................................1.0g
NH4Cl .....................................................................1.0g
L -Cysteine................................................................0.5g
Na-acetate ................................................................0.5g
K2HPO4 ....................................................................0.3g
KH2PO4 ....................................................................0.3g
CaCl2 ·2H2O...............................................................0.1g
KCl........................................................................0.1g
Resazurin ................................................................0.5mg
NaHCO3  solution .........................................................50.0mL
Na-thiosulfate solution...................................................20.0mL
Na2S·9H2O solution.......................................................15.0mL
Trace elements solution ..................................................10.0mL
pH 7.3 ± 0.2 at 25°C
Trace Elements Solution:
Composition per liter:
MgSO4 ·7H2O.............................................................3.0g
Nitrilotriacetic acid....................................................1.5g
NaCl.....................................................................1.0g
MnSO4 ·2H2O.............................................................0.5g
CoSO4 ·7H2O............................................................0.18g
ZnSO4 ·7H2O............................................................0.18g
CaCl2 ·2H2O ............................................................0.1g
FeSO4 ·7H2O.............................................................0.1g
NiCl2 ·6H2O...........................................................0.025g
KAl(SO4)2 ·12H2O.......................................................0.02g
H3BO3 ..................................................................0.01g
Na2MoO4 ·4H2O..........................................................0.01g
CuSO4 ·5H2O............................................................0.01g
Na2SeO3 ·5H2O..........................................................0.3mg
Preparation of Trace Elements Solution: Add nitrilotriacetic
acid to 500.0mL of distilled/deionized water. Dissolve by adjusting pH
to 6.5 with KOH. Add remaining components. Add distilled/deionized
water to 1.0L. Mix thoroughly.
Na2S·9H2O Solution :
Composition per 20.0mL:
Na2S·9H2O..............................................................0.6g
Preparation of Na2S·9H2O Solution: Add Na2S·9H2O to dis-
tilled/deionized water and bring volume to 20.0mL. Mix thoroughly.
Autoclave under 100% N2 for 15 min at 15 psi pressure–121°C. Cool
to room temperature.
NaHCO3 Solution:
Composition per 100.0mL:
NaHCO3 .................................................................10.0g
Preparation of NaHCO3 Solution:  Add NaHCO3 to distilled/de-
ionized water and bring volume to 100.0mL. Mix thoroughly. Sparge
with 80% N2 + 20% CO2 . Filter sterilize.
Na-thiosulfate Solution:
Composition per 20.0mL:
Na2S2O3 ·5H2O.........................................................5.0g
Preparation of Na-thiosulfate Solution: Add Na2S2O3 ·5H2O to
distilled/deionized water and bring volume to 20.0mL. Mix thorough-
ly. Sparge with 100% N2 . Filter sterilize.
Preparation of Medium: Prepare and dispense medium under 80%
N2 + 20% CO2.  Add components, except Na-thiosulfate solution,
NaHCO3  solution, and Na2S·9H2O solution, to distilled/deionized wa-
ter and bring volume to 1.0L. Mix thoroughly. Sparge with 80% N2 +
20% CO2 . Autoclave for 15 min at 15 psi pressure–121°C. Cool to
25°C. Aseptically and anaerobically add 20.0mL Na-thiosulfate solu-
tion, 50.0mL NaHCO3  solution, and 15.0mL Na2S·9H2O solution. Mix
thoroughly. Adjust pH to 7.3. Aseptically and anaerobically distribute
into sterile tubes or flasks.
Use:  For the cultivation of Dethiosulfovibrio peptidovorans.