Desulfohalobium utahense Medium with Malate (DSMZ Medium 1055)

[所属分类:培养基配方] [发布时间:2020-6-8] [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

Desulfohalobium utahense Medium with Malate
(DSMZ Medium 1055)

培养基配方
Composition per liter:
NaCl .......................................................................10.0g
MgSO4 ·7H2O................................................................10.0g
KCl..........................................................................6.0g
NH4Cl .......................................................................1.0g
KH2PO4 ......................................................................0.1g
CaCl2 ·2H2O ................................................................0.4g
Yeast extract................................................................0.5g
Resazurin ..................................................................0.5mg
NaHCO3  solution...........................................................10.0mL
Malate solution ...........................................................10.0mL
Na2S·9H2O solution........................................................10.0mL
Trace elements solution SL-10...............................................1.0mL
Selenite/tungstate solution.................................................1.0mL
pH 7.1 ± 0.2 at 25°C
Malate Solution :
Composition per 10.0mL:
Na2( DL )-malate .............................................................1.0g
Preparation of Malate Solution: Add Na2 ( DL )-malate to distilled/
deionized water and bring volume to 10.0mL. Mix thoroughly. Auto-
clave under 100% N2 for 15 min at 15 psi pressure–121°C. Cool to
room temperature.
Na2S·9H2O Solution :
Composition per 10.0mL:
Na2S·9H2O.....................................................................0.1g
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 10.0mL:
NaHCO3 .......................................................................4.0g
Preparation of NaHCO3 Solution:  Add NaHCO3 to distilled/de-
ionized water and bring volume to 10.0mL. Mix thoroughly. Sparge
with 20% CO2 + 80% H2 . Autoclave for 15 min at 15 psi pressure–
121°C. Cool to room temperature.
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 . Autoclave for 15 min at 15 psi pressure–121°C.
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 100% N2 . Autoclave for 15 min at 15 psi pressure–
121°C.
Preparation of Medium:  Add components, except bicarbonate, vi-
tamins, and reducing agents, to distilled/deionized water and bring vol-
ume to 960.0mL. Adjust pH to 7.2. Gently heat and bring to boiling.
Boil for 1 min. Mix thoroughly. Cool to room temperature while sparg-
ing with 80% N2  + 20% CO2 . Dispense into culture vessels under an
atmosphere of 80% N2  + 20% CO2 . Autoclave for 15 min at 15 psi pres-
sure–121°C. Aseptically add bicarbonate, sodium lactate, and sulfide.
Adjust the final pH of the medium to 7.0–7.2.
Use: For the cultivation of Desulfovermiculus halophilus.