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山東拓普生物工程有限公司

Shandong Tuopu Biol-Engineering Co.,Ltd

Eubacterium acidaminophilum Medium

培養(yǎng)基配方
Composition per liter:
KH2PO4 .....................................................................1.0g
NH4Cl ......................................................................0.5g
MgCl2 ·6H2O ...............................................................0.4g
CaCl2 ·2H2O ...............................................................0.1g
NaHCO3  solution..........................................................20.0mL
Sodium selenite solution..................................................10.0mL
Na2S·9H2O solution........................................................3.0mL
Vitamin solution...........................................................1.0mL
Trace elements solution SL-7...............................................1.0mL
pH 7.2–7.3 at 25°C
Sodium Selenite Solution:
Composition per liter:
NaOH........................................................................0.5g
Na2SeO3 ·5H2O.............................................................0.03g
Preparation of Sodium Selenite Solution: Add components to
distilled/deionized water and bring volume to 1.0L. Mix thoroughly.
Filter sterilize. Gas with 100% N2 for 20 min.
Na2S·9H2O Solution:
Composition per 10.0mL:
Na2S·9H2O.............................................................1.0g
Preparation of Na2S·9H2O Solution: Add Na2S·9H2O to dis-
tilled/deionized water and bring volume to 10.0mL. Mix thoroughly.
Gas with 100% N2 for 20 min. Autoclave for 15 min at 15 psi pressure–
121°C. Cool to 45°–50°C.
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 100.0mL:
p-Aminobenzoic acid......................................................4.0mg
Biotin...................................................................1.0mg
Preparation of Vitamin Solution: Add components to distilled/
deionized water and bring volume to 100.0mL. Mix thoroughly. Filter
sterilize.
Trace Elements Solution SL-7:
Composition per liter:
FeCl2 ·4H2O..............................................................1.5g
CoCl2 ·6H2O ............................................................0.19g
MnCl2 ·4H2O..............................................................0.1g
ZnCl2 ...................................................................0.07g
Na2MoO4 ·2H2O .........................................................0.036g
NiCl2 ·6H2O............................................................0.024g
H3BO3 ..................................................................6.0mg
CuCl2 ·2H2O ...........................................................2.0mg
HCl (25% solution).....................................................10.0mL
Preparation of Trace Elements Solution SL-7: Add the
FeCl2 ·4H2O to the HCl. Add distilled/deionized water and bring volume
to 1.0L. Add remaining components. Mix thoroughly. Autoclave for 15
min at 15 psi pressure–121°C under 100% N2 . Cool to 25°C.
Preparation of Medium: Add components—except NaHCO3  solu-
tion, Na2S·9H2O solution, vitamin solution, and sodium selenite solu-
tion—to distilled/deionized water and bring volume to 966.0mL. Mix
thoroughly. Gently heat and bring to boiling. Autoclave for 15 min at
15 psi pressure–121°C. Cool to 45°–50°C under 80% N2 and 20% CO2 .
Aseptically add 20.0mL of sterile NaHCO3  solution, 3.0mL of sterile
Na2S·9H2O solution, 1.0mL of sterile vitamin solution, and 10.0mL of
sterile sodium selenite solution. Mix thoroughly. Adjust pH to 7.2–7.3
with dilute sterile HCl or Na2CO3 , if necessary. Aseptically and anaero-
bically distribute into sterile tubes under 80% N2 and 20% CO2 . Cap
tubes with rubber stoppers.
Use: For the cultivation and maintenance of Eubacterium acidamino-
philum.