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BS Medium
山东拓普生物工程有限公司
Shandong Tuopu Biol-Engineering Co.,Ltd
BS Medium
Composition per 1135.0mL:
NaHCO3 ...............................................................2.2g
NH4Cl ...............................................................0.25g
KH2PO4 ..............................................................0.07g
Resazurin............................................................0.5mg
(NH4)2(Fe(SO4)2 ·6H2O..............................................0.2mg
Na2SeO4 .............................................................0.1mg
Na2WO4 ·2H2O........................................................0.1mg
Marine medium/synthetic seawater mix................................125.0mL
Wolfe’s mineral solution...........................................10.0mL
Yeast extract solution..............................................10.0mL
KNO3 solution......................................................10.0mL
pH 7.0 ± 0.2 at 25°C
Marine Medium/Synthetic Seawater Mix:
Composition per liter:
NaCl.................................................................47.15g
MgCl2 ·6H2O ........................................................18.1g
MgSO4 ·7H2O.........................................................7.0g
Na2SO4 .............................................................3.24g
CaCl2 ·2H2O .......................................................3.13g
KCl..................................................................1.2g
Na2CO3 ..............................................................0.1g
NaBr.................................................................0.1g
KBr ...............................................................80.0mg
SrCl2 ·6H2O ......................................................72.0mg
H3BO3 .............................................................52.0mg
Na2HPO4 ...........................................................8.1mg
NaF ...............................................................2.4mg
Sodium silicate....................................................0.4mg
KI................................................................50.0μg
Preparation of Marine Medium/Synthetic Seawater Mix:
Add components to distilled/deionized water and bring volume to 1.0L.
Mix thoroughly.
Wolfe’s Mineral Solution:
Composition per liter:
MgSO4 ·7H2O..................................................3.0g
Nitrilotriacetic acid.........................................1.5g
NaCl..........................................................1.0g
MnSO4 ·2H2O..................................................0.5g
CoCl2 ·6H2O..................................................0.1g
ZnSO4 ·7H2O..................................................0.1g
CaCl2 ·2H2O .................................................0.1g
FeSO4 ·7H2O..................................................0.1g
NiCl2 ·6H2O................................................0.025g
KAl(SO4)2 ·12H2O............................................0.02g
CuSO4 ·5H2O.................................................0.01g
H3BO3 .......................................................0.01g
Na2MoO4 ·2H2O...............................................0.01g
Na2SeO3 ·5H2O...............................................0.3mg
Preparation of Wolfe’s Mineral Solution: Add nitrilotriacetic
acid to 500.0mL of distilled/deionized water. Adjust pH to 6.5 with
KOH. Add remaining components. Add distilled/deionized water to
1.0L. Adjust pH to 6.8.
Yeast Extract Solution:
Composition per 10.0mL:
Yeast extract................................................0.5g
Preparation of Yeast Extract Solution: Add yeast extract to dis-
tilled/deionized water and bring volume to 10.0mL. Mix thoroughly.
Sparge with 80% N 2 + 20% CO 2 . Autoclave for 15 min at 15 psi pres-
sure–121°C.
KNO3 Solution:
Composition per 10.0mL:
KNO3 ...........................................................1.0g
Preparation of KNO 3 Solution: Add KNO 3 to distilled/deionized
water and bring volume to 10.0mL. Mix thoroughly. Sparge with 80%
N2 + 20% CO2 . Autoclave for 15 min at 15 psi pressure–121°C.
Preparation of Medium: Add components, except yeast extract so-
lution and KNO3 solution, to distilled/deionized water and bring vol-
ume to 980.0mL. Mix thoroughly. Adjust pH to 7.0 with H2SO4 .
Distribute 20.0mL volumes into 100.0mL bottles. Sparge with 80% N2
+ 20% CO2 . Autoclave for 15 min at 15 psi pressure–121°C. Aseptical-
ly and anaerobically add 0.2mL of sterile yeast extract solution and
0.2mL of sterile KNO3 solution to each bottle. After inoculation, pres-
surize bottles to 2 bar with 80% N2 + 20% CO2 .
Use: For the cultivation of Pyrobaculum aerophilum.