Acetitomaculum Medium

[所属分类:培养基配方] [发布时间:2019-9-18] [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

Acetitomaculum Medium
Composition per liter:
NaCl..............................................................................................7.0g
Glucose.........................................................................................5.0g
NaHCO 3 ........................................................................................4.0g
Yeast extract..................................................................................1.5g
MgCl 2 ·6H 2 O .................................................................................1.2g
KCl................................................................................................0.5g
NH 4 Cl ...........................................................................................0.3g
CaCl 2 ·2H 2 O ................................................................................0.15g
Na 2 SO 4 ..........................................................................................0.1g
NiCl 2 ·6H 2 O................................................................................1.0mg
Resazurin...................................................................................0.5mg
Na 2 WO 4 ·2H 2 O...........................................................................0.1mg
Ascorbic acid............................................................................50.0μg
Choline chloride........................................................................50.0μg
D -myo-Inositol..........................................................................50.0μg
Nicotinamide.............................................................................50.0μg
Glucose solution......................................................................50.0mL
Trace elements solution...........................................................10.0mL
Vitamin solution.......................................................................10.0mL
Reducing agent solution ..........................................................10.0mL
pH 7.0 ± 0.2 at 25°C
Glucose Solution:
Composition per 50.0mL:
D- Glucose......................................................................................5.0g
Preparation of Glucose Solution: Add glucose to distilled/deion-
ized water and bring volume to 50.0mL. Mix thoroughly. Dispense so-
lution anaerobically under 100% N 2 gas. Autoclave for 15 min at 15 psi
pressure–121°C.
Trace Elements Solution:
Composition per liter:
MgSO 4 ·7H 2 O................................................................................3.0g
Nitrilotriacetic acid.......................................................................1.5g
CaCl 2 ·2H 2 O..................................................................................1.0g
NaCl..............................................................................................1.0g
MnSO 4 ·2H 2 O...............................................................................0.5 g
CoSO 4 ·7H 2 O..............................................................................0.18 g
ZnSO 4 ·7H 2 O..............................................................................0.18 g
FeSO 4 ·7H 2 O.................................................................................0.1g
NiCl 2 ·6H 2 O..............................................................................0.025 g
KAl(SO 4 ) 2 ·12H 2 O......................................................................0.02g
CuSO 4 ·5H 2 O...............................................................................0.01g
H 3 BO 3 .........................................................................................0.01g
Na 2 MoO 4 ·2H 2 O..........................................................................0.01g
Na 2 SeO 3 ·5H 2 O..........................................................................0.3 mg
Preparation of Trace Elements Solution : Add nitrilotriacetic
acid to 500.0mL of distilled/deionized water. Dissolve by adjusting pH
to 6.5 with KOH. Add distilled/deionized water to 1.0L. Add remain-
ing components. Mix thoroughly.
Vitamin Solution:
Composition per liter:
Pyridoxine·HCl........................................................................10.0mg
Calcium  DL -pantothenate...........................................................5.0mg
Lipoic acid.................................................................................5.0mg
Nicotinic acid.............................................................................5.0mg
p-Aminobenzoic acid.................................................................5.0mg
Riboflavin..................................................................................5.0mg
Thiamine·HCl............................................................................5.0mg
Biotin.........................................................................................2.0mg
© 2010 by Taylor and Francis Group, LLC
Folic acid....................................................................................2.0mg
Vitamin B 12 ................................................................................0.1mg
Preparation of Vitamin Solution: Add components to distilled/
deionized water and bring volume to 1.0L. Mix thoroughly.
Reducing Agent Solution:
Composition per 110.0mL:
L -Cysteine·HCl·H 2 O .....................................................................2.5g
Na 2 S·9H 2 O....................................................................................2.5g
Preparation of Reducing Agent Solution : Add 110.0mL of dis-
tilled/deionized water to a 250.0mL flask. Boil under N 2 gas for 1 min. Cool
to room temperature. Add  L -cysteine·HCl·H 2 O and dissolve. Adjust to pH 9
with 5N NaOH. Add washed Na 2 S·9H 2 O and dissolve. Distribute in
10.0mL volumes into tubes. Autoclave for 10 min at 15 psi pressure–121°C.
Preparation of Medium: Add components, except NaHCO 3 , glu-
cose solution, and reducing agent solution, to distilled/deionized water
and bring volume to 920.0mL. Gently heat and bring to boiling. Con-
tinue boiling for 3 min. Cool to room temperature under 80% N 2 + 20%
CO 2 . Add solid NaHCO 3 and bring pH to 6.8–7.0 by gassing. Distrib-
ute anaerobically under 80% N 2 + 20% CO 2 . Autoclave for 15 min at
15 psi pressure–121°C. Prior to inoculation of cultures, aseptically and
anaerobically add 0.1mL of sterile reducing agent solution and 0.5mL
of sterile glucose solution to each tube containing 9.4mL of sterile bas-
al medium.
Use: For the cultivation and maintenance of Acetitomaculum ruminus.