Bacillus selenitireducens Medium (DSMZ Medium 968)

[所属分类:培养基配方] [发布时间:2019-12-2] [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

Bacillus selenitireducens Medium
(DSMZ Medium 968）

培养基配方
Composition per liter:
NaCl.............................................................90.0g
Na2CO3 ..........................................................10.6g
NaHCO3 ...........................................................4.2g
Na-lactate.......................................................1.70g
NaNO3 ...........................................................1.25g
Yeast extract.....................................................0.2g
K2HPO4 ..........................................................0.15g
(NH4)SO4 .........................................................0.1g
KH2PO4 ..........................................................0.08g
MgSO4 ..........................................................25.0mg
Resazurin.......................................................0.5mg
Cysteine solution................................................10.0mL
Na2S·9H2O solution..............................................10.0mL
Trace elements solution SL-10 ....................................1.0mL
Selenite tungstate solution.......................................1.0mL
pH 9.8 ± 0.2 at 25°C
Cysteine Solution:
Composition per 10.0mL:
L -Cysteine·HCl·H2O .............................................0.25g
Preparation of Cysteine Solution:  Add  L -cysteine·HCl·H 2 O to
distilled/deionized water and bring volume to 10.0mL. Mix thorough-
ly. Sparge with 100% N2 . Autoclave for 15 min at 15 psi pressure–
121°C.
Na2S·9H2O Solution:
Composition per 10.0mL:
Na2S·9H2O.......................................................0.25g
Preparation of Na 2 S·9H2O Solution:  Add Na2S·9H2O to dis-
tilled/deionized water and bring volume to 10.0mL. Sparge with N 2 .
Autoclave for 15 min at 15 psi pressure–121°C. Cool to 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.
Selenite Tungstate Solution
Composition per liter:
NaOH.................................................................0.5g
Na2WO4 ·2H2O.......................................................4.0mg
Na2SeO3 ·5H2O......................................................3.0mg
Preparation of Selenite Tungstate Solution: Add components
to distilled/deionized water and bring volume to 1.0L. Mix thoroughly.
Sparge with 100% N2 . Filter sterilize.
Preparation of Medium: Add components, except NaHCO3 ,
Na2CO3 , cysteine solution, and Na2S·9H2O solution, to distilled/deion-
ized water and bring volume to 980.0mL. Mix thoroughly. Gently heat
and bring to boiling. Boil for 3 min. Cool to room temperature while
sparging with 80% N2  gas. Add solid NaHCO3  and Na2CO3 . Adjust pH
to 9.8. Distribute to anaerobe tubes or bottles. Autoclave for 15 min at
15 psi pressure–121°C. Cool to room temperature. Aseptically and an-
aerobically add per liter 10.0mL sterile cysteine solution and 10.0mL
sterile Na2S·9H2O solution. Mix thoroughly.
Use:  For the cultivation of Bacillus selenitireducens and Bacillus
arseniciselenatis.