ARC51 Medium (DSMZ Medium 1098)

[所属分类:培养基配方] [发布时间:2019-11-6] [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

ARC51 Medium
(DSMZ Medium 1098)
Composition per liter:
Sea salts, Sigma..........................................................................35.0g
Sodium acetate .............................................................................1.6g
Mercaptoethanesulfonic acid (coenzyme M) .............................0.14g
NH4Cl ...........................................................................................0.1g
Yeast extract..................................................................................0.1g
K2HPO4 .......................................................................................0.05g
Resazurin ..................................................................................0.5mg
NaHCO3  solution .....................................................................10.0mL
Na2S·9H2O solution.................................................................10.0mL
Na2S2O3  solution......................................................................10.0mL
Wolfe’s mineral elixer............................................................... 1.0mL
Seven vitamin solution...............................................................1.0mL
pH 6.5 ± 0.2 at 25°C
Seven Vitamin Solution:
Composition per liter:
Pyridoxine hydrochloride ......................................................300.0mg
Thiamine-HCl·2H 2 O..............................................................200.0mg
Nicotinic acid.........................................................................200.0mg
Vitamin B 12 ............................................................................100.0mg
Calcium pantothenate ............................................................100.0mg
p-Aminobenzoic acid...............................................................80.0mg
D (+)-Biotin...............................................................................20.0mg
Preparation of Seven Vitamin Solution: Add components to dis-
tilled/deionized water and bring volume to 1.0L. Sparge with 100% N 2 .
Mix thoroughly. Filter sterilize.
Wolfe’s Mineral Elixir:
Composition per liter :
MgSO4 ·7H2O..............................................................................30.0g
NaCl............................................................................................10.0g
MnSO4 ·2H2O................................................................................5.0g
(NH4)2NiSO4 ·6H2O ......................................................................2.8g
CoCl2 ·6H2O ..................................................................................1.8g
ZnSO4 ·7H2O.................................................................................1.8g
FeSO4 ·7H20..................................................................................1.0g
CaCl2 ·2H2O...................................................................................1.0g
KAl(SO4) 2 ·12H2O.......................................................................0.18g
CuSO4 ·5H2O.................................................................................0.1g
H3BO3 ...........................................................................................0.1g
Na2MoO4 ·2H2O ............................................................................0.1g
Na2SeO4 ........................................................................................0.1g
Na2WO4 ·2H2O..............................................................................0.1g
Preparation of Wolfe’s Mineral Elixir: Adjust pH of 1.0L of dis-
tilled/deionized water to 1.0 with dilute H 2 SO 4 . Add remaining com-
ponents one at a time. Mix thoroughly to dissolve.
Na2S·9H2O Solution :
Composition per 10.0mL:
Na2S·9H20....................................................................................0.5g
Preparation of Na 2 S·9H 2 O Solution: Add Na 2 S·9H 2 O to dis-
tilled/deionized water and bring volume to 10.0mL. Mix thoroughly.
Autoclave under 100% N 2 for 15 min at 15 psi pressure–121°C. Cool
to room temperature.
Na2S2O3 Solution :
Composition per 10.0mL:
Na2S2O3 ·5H2O..............................................................................2.5g
Preparation of NaHCO 3 Solution:  Add Na 2 S 2 O 3 ·5H 2 O to dis-
tilled/deionized water and bring volume to 10.0mL. Mix thoroughly.
Sparge with N 2 . Autoclave for 15 min at 15 psi pressure–121°C. Cool
to room temperature.
NaHCO3 Solution :
Composition per 50.0mL:
NaHCO3 ........................................................................................2.0g
Preparation of NaHCO 3 Solution:  Add NaHCO 3 to distilled/de-
ionized water and bring volume to 50.0mL. Mix thoroughly. Sparge
with N 2 . Autoclave for 15 min at 15 psi pressure–121°C. Cool to room
temperature.
Preparation of Medium: Add components, except vitamin solu-
tion, bicarbonate solution, thiosulfate solution, and Na 2 S·9H 2 O solu-
tion, to distilled/deionized water and bring volume to 970.0mL. Mix
thoroughly. Gently heat and bring to boiling. Boil for 3 min. Cool to
room temperature while sparging with 20% CO 2 + 80% N 2 . Dispense
into balch tubes or serum bottles under atmosphere of 20% CO 2 + 80%
N 2 . Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C under
an atmosphere of 20% CO 2 + 80% N 2 . Aseptically and anaerobically
add sterile vitamin solution, bicarbonate solution, thiosulfate solution,
and Na 2 S·9H 2 O solution. Adjust final pH to 6.5. After inoculation ad-
just overpressure to 1.5 bar with 20% CO 2 + 80% N 2 .
Use: For the cultivation and maintenance of Archaeoglobus infectus.