​CP Medium for Thermobacteroides leptospartum

[所属分类:培养基配方] [发布时间:2020-5-26] [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

CP Medium for Thermobacteroides leptospartum

培养基配方
Composition per 1010.0mL:
NaHCO3 ....................................................................8.4g
Pancreatic digest of casein................................................2.0g
Yeast extract..............................................................2.0g
MgCl2 ·6H2O...............................................................1.0g
NH4Cl .....................................................................1.0g
CaCl2 ·2H2O...............................................................0.4g
K2HPO4 ·3H2O..............................................................0.4g
Resazurin ................................................................0.5mg
Glucose solution .......................................................100.0mL
Na2S·9H2O solution......................................................10.0mL
Wolfe’s mineral solution................................................10.0mL
pH 7.0 ± 0.2 at 25°C
Glucose Solution:
Composition per 100.0mL:
D -Glucose.................................................................5.0g
Preparation of Glucose Solution:  Add glucose to distilled/deion-
ized water and bring volume to 100.0mL. Mix thoroughly. Sparge with
100% N2 . Autoclave for 15 min at 15 psi pressure–121°C.
Na2S·9H2O Solution:
Composition per 10.0mL:
Na2S·9H2O.................................................................0.5g
Preparation of Na2S·9H2O Solution:  Add Na2S·9H2O to dis-
tilled/deionized water and bring volume to 10.0mL. Mix thoroughly.
Sparge with 100% N2 . Autoclave for 15 min at 15 psi pressure–121°C.
Before use, neutralize to pH 7.0 with sterile HCl.
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.
Preparation of Medium: Prepare medium anaerobically under 100%
CO2 . Add components, except glucose solution, Na2S·9H2O solution, and
Wolfe’s mineral solution, to distilled/deionized water and bring volume to
880.0mL. Mix thoroughly. Gently heat and bring to boiling. Cool to room
temperature while sparging with 100% CO2 . Sparge with 100% CO2 for
20 min. Adjust pH to 7.0. Autoclave for 15 min at 15 psi pressure–121°C.
Aseptically and anaerobically add 100.0mL of sterile glucose solution,
10.0mL of sterile Na2S·9H2O solution, and 10.0mL of sterile Wolfe’s min-
eral solution to each tube. Mix thoroughly.
Use:  For the cultivation of Thermobacteroides leptospartum.