Benzoate Nitrate Salts Medium (BNS)

[所属分类:培养基配方] [发布时间:2019-12-16] [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　

Benzoate Nitrate Salts Medium
(BNS)

培养基配方
Composition per liter:
Solution A...................................................700.0mL
Solution B...................................................300.0mL
pH 8.2 ± 0.2 at 25°C
Solution A:
Composition per 700.0mL:
KNO3 .........................................................2.0g
Sodium benzoate...............................................1.0g
NH4Cl ........................................................0.3g
Phosphate buffer solution..................................200.0mL
Phosphate Buffer Solution:
Composition per 200.0mL:
K2HPO4 .....................................................5.12g
KH2PO4 ......................................................1.5g
Preparation of Phosphate Buffer: Add components to distilled/de-
ionized water and bring volume to 200.0mL. Mix thoroughly. Adjust pH
to 9.0 with KOH.
Preparation of Solution A:  Add components to distilled/deionized
water and bring volume to 700.0mL. Mix thoroughly. Autoclave for 15
min at 15 psi pressure–121°C. Cool to room temperature.
Solution B:
Composition per 300.0mL:
MgSO4 ·7H2O.................................................0.2g
CaCl2 .....................................................10.0mg
Trace metals solution.......................................1.0mL
Preparation of Solution B:  Add components to distilled/deionized
water and bring volume to 300.0mL. Mix thoroughly. Autoclave for 15
min at 15 psi pressure–121°C. Cool to room temperature.
Trace Metals Solution:
Composition per 300.0mL:
MnSO4 ·H2O................................................50.0mg
ZnSO4 ·7H2O...............................................50.0mg
Co(NO3)2 ·6H2O............................................10.0mg
CuSO4 .....................................................10.0mg
Na2B4O7 ·10H2O............................................10.0mg
Na2MoO4 ·2H2O .............................................0.2mg
Ferric EDTA solution.......................................10.0mL
Preparation of Trace Metals Solution: Add components to dis-
tilled/deionized water and bring volume to 1.0L. Mix thoroughly.
Ferric EDTA Solution:
Composition per 550.0mL:
EDTA........................................................17.9g
FeSO4 ·7H2O................................................13.7g
KOH.........................................................3.23g
Preparation of Ferric EDTA Solution: Add EDTA and KOH to
distilled/deionized water and bring volume to 186.0mL. Mix thorough-
ly. In a separate flask, add FeSO4 ·7H2O to distilled/deionized water
and bring volume to 364.0mL. Mix thoroughly. Combine the two solu-
tions. Sparge with air overnight to oxidize the Fe 2+ to Fe 3 + . Store in the
dark.
Preparation of Medium: Aseptically combine 700.0mL of sterile
solution A with 300.0mL of sterile solution B. Adjust pH to 8.2. Asepti-
cally distribute into sterile screw-capped tubes. Fill tubes completely.
Use: For the cultivation of Alcaligenes xylosoxydans.