Halophilic Clostridium Broth

[所属分类:培养基配方] [发布时间:2021-3-30] [发布人: ] [阅读次数:] [返回]

Halophilic Clostridium Broth

培养基配方

Composition per liter:
L- Cysteine·HCl·H 2 O .....................................................................0.5g
Solution 1......................................................................................1.0L
Solution 2...............................................................................100.0mL
pH 6.2–7.0 at 25°C
Solution 1:
Composition per liter:
NaCl..........................................................................................105.0g
KCl................................................................................................7.5g
L- Glutamic acid.............................................................................4.0g
Casamino acids.............................................................................2.0g
Nutrient broth................................................................................2.0g
Yeast extract..................................................................................2.0g
FeSO 4 ·7H 2 O...............................................................................2.0mg
Resazurin...................................................................................1.0mg
NaOH (2.5N solution)..............................................................12.5mL
Wolfe’s vitamin solution..........................................................10.0mL
Wolfe’s mineral solution..........................................................10.0mL
Preparation of Solution 1: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly.
Wolfe’s Vitamin Solution:
Composition per liter:
Pyridoxine·HCl...........................................................................0.01g
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.
Wolfe’s Mineral Solution:
Composition per liter:
MgSO 4 ·7H 2 O................................................................................3.0g
Nitrilotriacetic acid.......................................................................1.5g
NaCl..............................................................................................1.0g
MnSO 4 ·H 2 O..................................................................................0.5g
FeSO 4 ·7H 2 O..................................................................................0.1g
CoCl 2 ·6H 2 O..................................................................................0.1g
CaCl 2 .............................................................................................0.1g
ZnSO 4 ·7H 2 O.................................................................................0.1g
CuSO 4 ·5H 2 O...............................................................................0.01g
AlK(SO 4 ) 2 ·12H 2 O.......................................................................0.01g
H 3 BO 3 .........................................................................................0.01g
Na 2 MoO 4 ·2H 2 O..........................................................................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 remaining components. Add distilled/deionized water to 1.0L.
Solution 2:
Composition per 100.0mL:
MgCl 2 ·6H 2 O ...............................................................................20.3g
CaCl 2 ·2H 2 O ................................................................................7.35g
Preparation of Solution 2: Add components to distilled/deionized water and bring volume to 100.0mL. Mix thoroughly. Gas with 100% N 2 for 20 min. Autoclave for 15 min at 15 psi pressure–121°C.
Preparation of Medium: Gently heat 1.0L of solution 1 and bring to boiling under 100% N 2 . Add the  L -cysteine·HCl·H 2 O. Continue boiling until resazurin turns colorless, indicating reduction. The volume of solution 1 should be about 900.0mL. Anaerobically distribute into tubes in 9.0mL volumes under 100% N 2 . Cap tubes with rubber stoppers. Place tubes in a press. Autoclave for 15 min at 15 psi pressure–121°C with fast exhaust. Cool to 25°C. Aseptically add 1.0mL of sterile solution 2 to each tube. Adjust pH to 6.2–7.0 if necessary with sterile O 2 -free NaOH or HCl.
Use:  For the cultivation and maintenance of Sporohalobacter lortetii.