[所屬分類:培養(yǎng)基配方] [發(fā)布時間:2020-7-9] [發(fā)布人:] [閱讀次數(shù):] [返回]

山東拓普生物工程有限公司

Shandong Tuopu Biol-Engineering Co.,Ltd

Eubacterium aggregans Medium
(DSMZ Medium 711a)

培養(yǎng)基配方
Composition per liter:
Yeast extract..............................................................1.0g
NH4Cl .....................................................................1.0g
NaCl.......................................................................0.6g
Cysteine-HCl·H2O..........................................................0.5g
K2HPO4 ....................................................................0.3g
KH2PO4 ....................................................................0.3g
Yeast extract..............................................................0.2g
MgCl2 ·6H2O..............................................................0.2g
CaCl2 ·2H2O..............................................................0.1g
KCl.......................................................................0.1g
Resazurin ...............................................................0.5mg
NaHCO3 solution.........................................................40.0mL
Fructose solution.......................................................10.0mL
Na2S·9H2O solution.....................................................10.0mL
Trace elements solution SL-10 ..........................................1.5mL
pH 7.1 ± 0.2 at 25°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 80% N2 + 20% CO2 . Autoclave for 15 min at 15
psi pressure–121°C.
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.
Fructose Solution:
Composition per 10.0mL:
Fructose..................................................................5.0g
Preparation of Fructose Solution: Add fructose to distilled/de-
ionized water and bring volume to 10.0mL. Mix thoroughly. Sparge
with 100% N2 . Autoclave for 15 min at 15 psi pressure–121°C. Cool to
room temperature.
Preparation of Medium: Prepare and dispense medium under 80%
N2 + 20% CO2 gas atmosphere. Add components, except NaHCO3  solu-
tion, Na2S·9H2O solution, fructose solution, and trace elements solu-
tion SL-10, to distilled/deionized water and bring volume to 938.5mL.
Mix thoroughly. Adjust pH to 7.1. Sparge with 80% N2 + 20% CO2 . Au-
toclave for 15 min at 15 psi pressure–121°C. Aseptically and anaerobi-
cally add 40.0mL NaHCO3  solution, 10.0mL Na2S·9H2O solution,
10.0mL fructose solution, and 1.5mL trace elements solution SL-10.
Mix thoroughly. Aseptically and anaerobically distribute into sterile
tubes or bottles.
Use:  For the cultivation of Clostridium methoxybenzovorans and
Eubacterium aggregans.