Cholesterol Medium

[所属分类:培养基配方] [发布时间:2020-4-23] [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

Cholesterol Medium

培养基配方
Composition per 1030.0mL:
Solution A.....................................................500.0mL
Solution B.....................................................500.0mL
Amino acid solution.............................................20.0mL
Vitamin solution................................................10.0mL
pH 6.8 ± 0.2 at 25°C
Solution A:
Composition per liter:
(NH4)2SO4 ........................................................5.0g
KH2PO4 ...........................................................1.0g
MgSO4 ·7H2O......................................................0.5g
CaCl2 ·2H2O......................................................0.1g
NaCl..............................................................0.1g
Wolfe’s mineral solution.......................................10.0mL
Preparation of Solution A: Add components to distilled/deionized
water and bring volume to 1.0L. Mix thoroughly. Autoclave for 15 min
at 15 psi pressure–121°C. Cool to 45°–50°C.
Wolfe’s Mineral Solution:
Composition per liter:
MgSO4 ·7H2O.....................................................3.0g
Nitrilotriacetic acid............................................1.5g
NaCl.............................................................1.0g
MnSO4 ·H2O......................................................0.5g
CaCl2 ...........................................................0.1g
CoCl2 ·6H2O ....................................................0.1g
FeSO4 ·7H2O.....................................................0.1g
ZnSO4 ·7H2O.....................................................0.1g
AlK(SO4)2 ·12H2O...............................................0.01g
CuSO4 ·5H2O....................................................0.01g
H3BO3 ..........................................................0.01g
Na2MoO4 ·2H2O .................................................0.01g
Preparation of Wolfe’s Mineral Solution: Add nitrilotriacetic
acid to 500.0mL of distilled/deionized water. Dissolve by adjusting pH
to 6.5 with KOH. Add distilled/deionized water to 1.0L. Add remain-
ing components.
Solution B:
Composition per liter:
Noble agar......................................................15.0g
Cholesterol......................................................2.0g
Tween™ 80........................................................1.0g
Yeast extract....................................................0.5g
Preparation of Solution B: Add components to distilled/deionized
water and bring volume to 1.0L. Mix thoroughly. Gently heat to boiling.
Autoclave for 15 min at 15 psi pressure–121°C. Cool to 45°–50°C.
Amino Acid Solution:
Composition per 100.0mL:
L- Histidine.....................................................0.5g
DL- Methionine...................................................0.1g
DL- Tryptophan...................................................0.1g
Preparation of Amino Acid Solution: Add components to dis-
tilled/deionized water and bring volume to 100.0mL. Filter sterilize.
Vitamin Solution:
Composition per liter:
myo-Inositol...................................................200.0mg
Calcium pantothenate ...........................................40.0mg
Niacin..........................................................40.0mg
Pyridoxine·HCl.................................................40.0mg
Thiamine........................................................40.0mg
p-Aminobenzoic acid.............................................20.0mg
Riboflavin......................................................20.0mg
Biotin........................................................200.0μg
Folic acid ...................................................200.0μg
Preparation of Vitamin Solution: Add components to distilled/
deionized water and bring volume to 1.0L. Filter sterilize.
Preparation of Medium:  Combine cooled, sterile solution A and
cooled, sterile solution B. Aseptically add filter-sterilized amino acid
solution and vitamin solution. Adjust pH to 6.8. Pour into sterile Petri
dishes or distribute into sterile tubes.
Use:  For the cultivation of ATCC strain 31384.