Halobacteroides acetoethylicus Medium

[所属分类:培养基配方] [发布时间:2021-3-26] [发布人: ] [阅读次数:] [返回]

Halobacteroides acetoethylicus Medium

培养基配方

Composition per liter:
NaCl..........................................................................................100.0g
Pancreatic digest of casein..........................................................10.0g
Yeast extract..................................................................................3.0g
K 2 HPO 4 .........................................................................................1.5g
NH 4 Cl ...........................................................................................0.9g
KH 2 PO 4 .......................................................................................0.75g
MgCl 2 ·6H 2 O .................................................................................0.2g
Glucose solution ......................................................................25.0mL
Na 2 S·9H 2 O (10% solution)......................................................10.0mL
Trace elements solution.............................................................9.0mL
Wolfe’s Vitamin solution...........................................................5.0mL
Resazurin (0.025% solution) .....................................................4.0mL
FeSO 4 ·7H 2 O (10% solution)....................................................0.03mL
pH 7.3 ± 0.2 at 25°C
Glucose Solution:
Composition per 100.0mL:
Glucose.......................................................................................20.0g
Preparation of Glucose Solution: Add glucose to distilled/deionized water and bring volume to 100.0mL. Mix thoroughly. Filter sterilize. Sparge with 100% N 2 .
Trace Elements Solution:
Composition per liter:
Nitrilotriacetic acid.....................................................................12.5g
NaCl..............................................................................................1.0g
FeCl 3 ·4H 2 O...................................................................................0.2g

MnCl 2 ·4H 2 O .................................................................................0.1g

CaCl 2 ·2H 2 O...................................................................................0.1g
ZnCl 2 .............................................................................................0.1g
CuCl 2 ...........................................................................................0.02g
Na 2 SeO 3 ......................................................................................0.02g
CoCl 2 ·6H 2 O ..............................................................................0.017g
H 3 BO 3 .........................................................................................0.01g
Na 2 MoO 4 ·2H 2 O ..........................................................................0.01g
Preparation of Trace Elements 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.
Wolfe’s Vitamin Solution:
Composition per liter:
Pyridoxine·HCl........................................................................10.0mg
Thiamine·HCl ............................................................................5.0mg
Riboflavin ..................................................................................5.0mg
Nicotinic acid.............................................................................5.0mg
Calcium  DL -pantothenate...........................................................5.0mg
p-Aminobenzoic acid.................................................................5.0mg
Thioctic acid ..............................................................................5.0mg
Biotin .........................................................................................2.0mg
Folic acid....................................................................................2.0mg
Cyanocobalamin .....................................................................100.0μg
Preparation of Wolfe’s Vitamin Solution: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. 

Preparation of Medium: Add components, except glucose solution, to distilled/deionized water and bring volume to 975.0mL. Mix thoroughly. Autoclave for 15 min at 15 psi pressure–121°C. While still hot, aseptically add 25.0mL of the sterile glucose solution under 97% N 2 + 3% H 2 . Adjust pH to 7.3 if necessary. Aseptically and anaerobically distribute into tubes. Cap with rubber stoppers.
Use:  For the cultivation and maintenance of Halobacteroides acetoethylicus.