Acetohalobium Medium

[所属分类:培养基配方] [发布时间:2019-9-18] [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

Acetohalobium Medium
Composition per 1025.0mL:
NaCl..........................................................................................150.0g
MgCl 2 ·6H 2 O .................................................................................4.0g
CaCl 2 ·2H 2 O ................................................................................0.33g
KCl..............................................................................................0.33g
KH 2 PO 4 .......................................................................................0.33g
NH 4 Cl .........................................................................................0.33g
Resazurin ...................................................................................1.0mg
Trace elements solution ...........................................................10.0mL
Vitamin solution.......................................................................10.0mL
Trimethylamine·HCl solution..................................................10.0mL
Yeast extract solution.................................................................5.0mL
NaHCO 3 solution.......................................................................5.0mL
Na 2 S·9H 2 O solution...................................................................5.0mL
pH 7.6 ± 0.2 at 25°C
Trace Elements Solution:
Composition per liter:
MgSO 4 ·7H 2 O................................................................................3.0g
Nitrilotriacetic acid......................................................................1.5 g
CaCl 2 ·2H 2 O..................................................................................1.0g
NaCl..............................................................................................1.0g
MnSO 4 ·2H 2 O................................................................................0.5g
CoSO 4 ·7H 2 O...............................................................................0.18g
ZnSO 4 ·7H 2 O...............................................................................0.18g
FeSO 4 ·7H 2 O .................................................................................0.1g
NiCl 2 ·6H 2 O...............................................................................0.025g
KAI(SO 4 ) 2 ·12H 2 O......................................................................0.02g
CuSO 4 ·5H 2 O...............................................................................0.01g
H 3 BO 3 .........................................................................................0.01g
Na 2 MoO 4 ·2H 2 O..........................................................................0.01g
Na 2 SeO 3 ·5H 2 O...........................................................................0.3mg
Preparation of Trace Elements 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. Mix thoroughly. Adjust pH to 7.0 with KOH.
Vitamin Solution:
Composition per liter:
Pyridoxine·HCl........................................................................10.0mg
Calcium  DL -pantothenate...........................................................5.0mg
Lipoic acid .................................................................................5.0mg
Nicotinic acid.............................................................................5.0mg
p-Aminobenzoic acid.................................................................5.0mg
Riboflavin ..................................................................................5.0mg
Thiamine·HCl ............................................................................5.0mg
Biotin .........................................................................................2.0mg
Folic acid....................................................................................2.0mg
Vitamin B 12 ................................................................................0.1mg
Preparation of Vitamin Solution:  Add components to distilled/
deionized water and bring volume to 1.0L. Mix thoroughly. Filter ster-
ilize.
Trimethylamine·HCl Solution:
Composition per 10.0mL:
Trimethylamine·HCl.....................................................................2.4g
Preparation of Trimethylamine·HCl Solution : Add trimethyl-
amine·HCl to distilled/deionized water and bring volume to 10.0mL.
Mix thoroughly. Sparge under 100% N 2 gas for 3 min. Autoclave for
15 min at 15 psi pressure–121°C. Store under N 2 gas. 4.5g of glycine
betaine may be used in place of trimethylamine·HCl.
Yeast Extract Solution:
Composition per 5.0mL:
Yeast extract................................................................................0.05g
Preparation of Yeast Extract Solution : Add yeast extract to dis-
tilled/deionized water and bring volume to 5.0mL. Mix thoroughly.
Sparge under 100% N 2 gas for 3 min. Autoclave for 15 min at 15 psi
pressure–121°C. Store under N 2 gas.
NaHCO 3 Solution:
Composition per 5.0mL:
NaHCO 3 ........................................................................................0.5g
Preparation of NaHCO 3 Solution : Add NaHCO 3 to distilled/de-
ionized water and bring volume to 5.0mL. Mix thoroughly. Sparge un-
der 100% N 2 gas for 3 min. Autoclave for 15 min at 15 psi pressure–
121°C. Store under N 2 gas.
Na 2 S·9H 2 O Solution:
Composition per 5.0mL:
Na 2 S·9H 2 O....................................................................................0.5g
Preparation of Na 2 S·9H 2 O Solution:  Add Na 2 S·9H 2 O to dis-
tilled/deionized water and bring volume to 5.0mL. Mix thoroughly.
Sparge under 100% N 2 gas for 3 min. Autoclave for 15 min at 15 psi
pressure–121°C. Store under N 2 gas.
Preparation of Medium:  Add components, except vitamin solu-
tion, trimethylamine·HCl solution, yeast extract solution, NaHCO 3 so-
lution, and Na 2 S·9H 2 O solution, to distilled/deionized water and bring
volume to 1.0L. Adjust pH to 7.2–7.4 with NaOH. Mix thoroughly.
Gently heat and bring to boiling. Boil for a few minutes. Allow to cool
to room temperature under 100% N 2 . Distribute into tubes or flasks un-
der 100% N 2 . Autoclave for 15 min at 15 psi pressure–121°C. Cool to
room temperature. Before inoculation, aseptically and anaerobically
add vitamin solution, trimethylamine·HCl solution, yeast extract solu-
tion, NaHCO 3 solution, and Na 2 S·9H 2 O solution. If necessary, adjust
pH to 7.6 with sterile anaerobic Na 2 CO 3 solution.
Use:  For the cultivation and maintenance of Acetohalobium arabati-
cum.