​Coal Medium

[所属分类:培养基配方] [发布时间:2020-5-20] [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

Coal Medium

培养基配方
Composition per 1011.0mL:
Coal, Pittsburgh seam .....................................................10.0g
NaHCO3 .....................................................................3.5g
Yeast extract...............................................................2.0g
NaCl........................................................................0.4g
NH4Cl ......................................................................0.4g
MgCl2 ·6H2O...............................................................0.33g
Na2S·9H2O..................................................................0.3g
CaCl2 ·2H2O...............................................................0.05g
Na2SeO3 ·5H2O............................................................3.0μg
KH2PO4 ....................................................................1.0mg
Resazurin .................................................................1.0mg
Wolfe’s vitamin solution..................................................10.0mL
Trace elements solution SL-10 ..............................................1.0mL
pH 7.3 ± 0.1 at 25°C
Wolfe’s Vitamin Solution:
Composition per liter:
Pyridoxine·HCl............................................................10.0mg
p-Aminobenzoic acid.........................................................5.0mg
Lipoic acid ................................................................5.0mg
Nicotinic acid..............................................................5.0mg
Riboflavin .................................................................5.0mg
Thiamine·HCl ..............................................................5.0mg
Calcium  DL -pantothenate...................................................5.0mg
Biotin .....................................................................2.0mg
Folic acid..................................................................2.0mg
Vitamin B 12 ...............................................................0.1mg
Preparation of Wolfe’s Vitamin Solution:  Add components to
distilled/deionized water and bring volume to 1.0L. Mix thoroughly.
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.
Preparation of Medium: Prepare and dispense medium under 80%
N2 + 20% CO2 . Add components, except NaHCO3 , to distilled/deion-
ized water and bring volume to 1.0L. Mix thoroughly. Gently heat and
bring to boiling. Continue boiling for 3 min. Cool to room temperature
while sparging with 80% N2 + 20% CO2 . Add NaHCO3 . Mix thorough-
ly. Adjust pH to 7.3. Anaerobically distribute 10.0mL volumes into an-
aerobic tubes. Autoclave for 15 min at 15 psi pressure–121°C. Adjust
pH to 7.3.
Use: For the cultivation of unidentified bacterium ATCC 55237.