EBA Medium

[所属分类:培养基配方] [发布时间:2020-7-2] [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

EBA Medium

培养基配方
Composition per 994.0mL:
Glycine......................................................................2.0g
NaCl.........................................................................1.0g
KCl..........................................................................0.5g
MgSO4 ·7H2O.................................................................0.5g
NH4Cl ......................................................................0.25g
KH2PO4 .....................................................................0.2g
CaCl2 ·2H2O ...............................................................0.15g
Resazurin..................................................................1.0mg
NaHCO3 solution............................................................30.0mL
Na2S·9H2O solution........................................................10.0mL
Biotin solution............................................................2.0mL
Na2SeO3 ·5H2O solution....................................................1.0mL
Trace elements solution SL-10 .............................................1.0mL
pH 7.2–7.4 at 25°C
NaHCO3 Solution:
Composition per 100.0mL:
NaHCO3 ...................................................................8.4g
Preparation of NaHCO3 Solution:  Add NaHCO3 to distilled/de-
ionized water and bring volume to 100.0mL. Mix thoroughly. Filter
sterilize. Gas under 80% N2 + 20% CO2 .
Na2S·9H2O Solution:
Composition per 10.0mL:
Na2S·9H2O................................................................0.6g
Preparation of Na2S·9H2O Solution:  Add Na2S·9H2O to dis-
tilled/deionized water and bring volume to 10.0mL. Mix thoroughly.
Gas under 100% N2 . Autoclave for 15 min at 15 psi pressure–121°C.
Biotin Solution:
Composition per 10.0mL:
Biotin ....................................................................0.1mg
Preparation of Biotin Solution:  Add biotin to distilled/deionized
water and bring volume to 10.0mL. Mix thoroughly. Gas under 100%
N2 . Autoclave for 10 min at 15 psi pressure–121°C.
Na2SeO3 ·5H2O Solution:
Composition per liter:
Na2SeO3 ·5H2O............................................................0.26g
NaOH (10mM solution)......................................................1.0L
Preparation of Na2SeO3 ·5H2O Solution : Add Na2SeO3 ·5H2O to
1.0L of 10mM NaOH solution. Mix thoroughly.
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. Gas under 100% N2 . Autoclave for 15 min at 15 psi pressure–
121°C.
Preparation of Medium: Add components, except NaHCO3 solu-
tion, Na2S·9H2O solution, and biotin solution, to distilled/deionized
water and bring volume to 952.0mL. Mix thoroughly. Sparge with
100% N2 . Autoclave for 15 min at 15 psi pressure–121°C. Aseptically
and anaerobically add 30.0mL of sterile NaHCO3 solution, 10.0mL of
sterile Na2S·9H2O solution, and 2.0mL of sterile biotin solution. Asep-
tically and anaerobically distibute into sterile flasks or tubes.
Use: For the cultivation and maintenance of Eubacterium acidamino-
philum.