Clostridium ultunense Medium (DSMZ Medium 727)

[所属分类:培养基配方] [发布时间:2020-5-18] [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

Clostridium ultunense Medium
(DSMZ Medium 727)

培养基配方
Composition per liter:
Yeast extract..............................................................10.0g
Peptone....................................................................10.0g
Lab Lemco powder............................................................5.0g
Na2HPO4 ...................................................................0.43g
Starch, soluble.............................................................0.4g
KH2PO4 .....................................................................0.4g
Na-acetate .................................................................0.4g
NH4Cl ......................................................................0.3g
NaCl........................................................................0.3g
CaCl2 ·2H2O................................................................0.1g
MgCl2 ·6H2O................................................................0.1g
Resazurin .................................................................0.5mg
NaHCO3 solution...........................................................50.0mL
Vitamin solution..........................................................10.0mL
Na2S·9H2O solution.......................................................10.0mL
Trace elements solution ...................................................1.0mL
Selenite-tungstate solution................................................1.0mL
pH 7.0 ± 0.2 at 25°C
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.
Na2S·9H2O Solution :
Composition per 10.0mL:
Na2S·9H2O.................................................................0.3g
Preparation of Na2S·9H2O Solution:  Add Na2S·9H2O to dis-
tilled/deionized water and bring volume to 10.0mL. 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.
Vitamin Solution:
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 Vitamin Solution:  Add components to distilled/
deionized water and bring volume to 1.0L. Mix thoroughly. Sparge
with 80% H2 + 20% CO2 . Filter sterilize.
Trace Elements Solution:
Composition per liter:
FeCl2 ·4H2O.............................................................1.5g
Na2 -EDTA................................................................0.5g
CoCl2 ·6H2O..........................................................190.0mg
MnCl2 ·4H2O .........................................................100.0mg
ZnCl2 .................................................................70.0mg
Na2MoO4 ·2H2O.........................................................36.0mg
NiCl2 ·6H2O...........................................................24.0mg
H3BO3 ..................................................................6.0mg
CuCl2 ·2H2O............................................................2.0mg
HCl (25% solution).....................................................10.0mL
Preparation of Trace Elements Solution: Add FeCl2 ·4H2O to
10.0mL of HCl solution. Mix thoroughly. Add distilled/deionized wa-
ter and bring volume to 1.0L. Add remaining components. Mix thor-
oughly. Sparge with 80% N2 + 20% CO2 . Autoclave for 15 min at 15
psi pressure–121°C.
Preparation of Medium: Prepare and dispense medium under 80%
N2 + 20% CO2 gas atmosphere. Add components, except NaHCO3  solu-
tion, Na2S·9H2O solution, vitamin solution, selenite-tungstate solution,
and trace elements solution, to distilled/deionized water and bring vol-
ume to 928.0mL. Mix thoroughly. Adjust pH to 7.0. Sparge with 80% N2
+ 20% CO2 . Autoclave for 15 min at 15 psi pressure–121°C. Aseptically
and anaerobically add 50.0mL NaHCO3  solution, 10.0mL Na2S·9H2O
solution, 10.0mL vitamin solution, 1.0mL selenite–tungstate solution,
and 1.0mL trace elements solution. Mix thoroughly. Aseptically and an-
aerobically distribute into sterile tubes or bottles.
Use:  For the cultivation of Clostridium ultunense.