Basal Thermophile Medium

[所属分类:培养基配方] [发布时间:2019-12-6] [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

Basal Thermophile Medium

培养基配方
Composition per liter:
Solution 1.................................................850.0mL
Solution 2.................................................100.0mL
Solution 3..................................................50.0mL
Solution 1:
Composition per 850.0mL:
Pancreatic digest of casein..................................10.0g
K2HPO4 .......................................................1.5g
NH4Cl ........................................................0.9g
KH2PO4 ......................................................0.75g
MgCl2 ·6H2O .................................................0.2g
Trace elements solution......................................9.0mL
Wolfe’s vitamin solution....................................5.0mL
Resazurin (0.2% solution) ...................................1.0mL
FeSO4 ·7H2O (10% solution).................................0.03mL
Preparation of Solution 1: Add components to distilled/deionized
water and bring volume to 850.0mL. Mix thoroughly. Autoclave for 45
min at 15 psi pressure–121°C. Cool to 45°–50°C.
Solution 2:
Composition per 100.0mL:
Yeast extract.................................................3.0g
Preparation of Solution 2:  Add yeast extract to distilled/deionized
water and bring volume to 100.0mL. Mix thoroughly. Autoclave for 45
min at 15 psi pressure–121°C. Cool to 45°–50°C.
Solution 3:
Composition per 50.0mL:
Glucose......................................................5.0g
Preparation of Solution 3: Add glucose to distilled/deionized wa-
ter and bring volume to 50.0mL. Mix thoroughly. Autoclave for 45 min
at 15 psi pressure–121°C. Cool to 45°–50°C.
Trace Elements Solution:
Composition per liter:
Nitrilotriacetic acid........................................12.5g
NaCl..........................................................1.0g
FeCl3 ·4H2O..................................................0.2g
MnCl2 ·4H2O .................................................0.1g
CaCl2 ·2H2O .................................................0.1g
ZnCl2 ........................................................0.1g
CuCl2 .......................................................0.02g
Na2SeO3 .....................................................0.02g
CoCl2 ·6H2O................................................0.017g
H3BO3 .......................................................0.01g
Na2MoO4 ·2H2O...............................................0.01g
Preparation of Trace Elements Solution:  Add nitrilotriacetic
acid to 100.0mL of distilled/deionized water. Adjust pH to 6.5 with
KOH. Add remaining components and bring volume to 1.0L. Mix thor-
oughly.
Wolfe’s Vitamin Solution:
Composition per liter:
Pyridoxine·HCl............................................10.0mg
Thiamine·HCl ..............................................5.0mg
Riboflavin .................................................5.0mg
Nicotinic acid..............................................5.0mg
Calcium pantothenate .......................................5.0mg
p-Aminobenzoic acid.........................................5.0mg
Thioctic acid ..............................................5.0mg
Biotin .....................................................2.0mg
Folic acid..................................................2.0mg
Cyanocobalamin .............................................0.1mg
Preparation of Wolfe’s Vitamin Solution: Add components to
distilled/deionized water and bring volume to 1.0L. Mix thoroughly.
Na2S·9H2O Solution:
Composition per 100.0mL:
Na2S·9H2O..................................................10.0g
Preparation of Na2S·9H2O Solution: Add Na2S·9H2O to dis-
tilled/deionized water and bring volume to 100.0mL. Mix thoroughly.
Autoclave for 15 min at 15 psi pressure–121°C.
Preparation of Medium:  Aseptically combine solution 1, solution
2, and solution 3 under 100% N 2 . Distribute into tubes in 10.0mL vol-
umes under 100% N2 . Immediately prior to inoculation, aseptically add
0.1mL of sterile Na2S·9H2O solution to each tube.
Use:  For the cultivation and maintenance of Clostridium species, Fer-
vidobacterium nodosum, and Thermoanaerobium brockii.