Clostridium bryantii Medium

[所属分类:培养基配方] [发布时间:2020-5-8] [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 bryantii Medium

培养基配方
Composition per liter:
NaCl....................................................................21.0g
MgCl2 ·6H2O ............................................................3.1g
KCl......................................................................0.5g
NH4Cl ...................................................................0.3g
KH2PO4 ..................................................................0.2g
CaCl2 ·2H2O ...........................................................0.15g
Resazurin...............................................................1.0mg
NaHCO3  solution.......................................................20.0mL
Na2S·9H2O solution....................................................20.0mL
Sodium caproate solution ..............................................20.0mL
Vitamin solution.......................................................20.0mL
Trace elements solution SL-10...........................................1.0mL
pH 7.2 ± 0.2 at 25°C
NaHCO3 Solution:
Composition per 20.0mL:
NaHCO3 ...................................................................5.0g
Preparation of NaHCO3 Solution:  Add NaHCO3 to distilled/de-
ionized water and bring volume to 20.0mL. Mix thoroughly. Sparge
with 80% N2 + 20% CO2 . Autoclave for 15 min at 15 psi pressure–
121°C.
Na2S·9H2O Solution:
Composition per 20.0mL:
Na2S·9H2O..................................................................0.4g
Preparation of Na2S·9H2O Solution:  Add Na2S·9H2O to dis-
tilled/deionized water and bring volume to 20.0mL. Mix thoroughly.
Sparge with 80% N2 + 20% CO2 . Autoclave for 15 min at 15 psi pres-
sure–121°C.
Sodium Caproate Solution:
Composition per 20.0mL:
Sodium caproate ..............................................................1.4g
Preparation of Sodium Caproate Solution:  Add sodium
caproate to distilled/deionized water and bring volume to 20.0mL. Mix
thoroughly. Sparge with 80% N2 + 20% CO2 . Autoclave for 15 min at
15 psi pressure–121°C.
Vitamin Solution:
Composition per 20.0mL:
Thiamine·HCl .............................................................100.0μg
p-Aminobenzoic acid.........................................................40.0μg
D (+)-Biotin................................................................10.0μg
Preparation of Vitamin Solution:  Add components to distilled/
deionized water and bring volume to 20.0mL. Mix thoroughly. Filter
sterilize. Sparge with 80% N2 + 20% CO2 .
Trace Elements Solution SL-10:
Composition per liter:
FeCl2 ·4H2O.................................................................1.5g
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 SL-10: Add FeCl2 ·4H2O
to 10.0mL of HCl solution. Mix thoroughly. Add distilled/deionized
water 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:  Add components, except NaHCO3  solution,
Na2S·9H2O solution, sodium caproate solution, and trace elements solu-
tion SL-10, to distilled/deionized water and bring volume to 919.0mL.
Mix thoroughly. Sparge with 80% N2 + 20% CO2 . Autoclave for 15 min
at 15 psi pressure–121°C. Aseptically and anaerobically add 20.0mL of
sterile NaHCO3  solution, 20.0mL of sterile Na2S·9H2O solution, 20.0mL
of sterile sodium caproate solution, and 1.0mL of sterile trace elements so-
lution SL-10. Mix thoroughly. Aseptically and anaerobically distribute
into sterile screw-capped bottles.
Use: For the cultivation and maintenance of Syntrophospora bryantii.