Low Phosphate Buffered Basal Medium, Modified

[所属分类:培养基配方] [发布时间:2021-5-8] [发布人:网站管理员2] [阅读次数:] [返回]

Low Phosphate Buffered Basal Medium, Modified

山东拓普生物工程有限公司 培养基配方 http://www.topbiol.com

Composition per 1030.0mL:

Pectin............................................................................................4.0g
NH 4 Cl ...........................................................................................1.0g
Na 2 HPO 4 .....................................................................................0.72g
KH 2 PO 4 .........................................................................................0.3g
MgCl 2 ·6H 2 O .................................................................................0.2g
Reducing agent ........................................................................20.0mL
Yeast extract solution...............................................................10.0mL
Trace minerals..........................................................................10.0mL
Wolfe’s vitamin solution............................................................5.0mL
Resazurin (0.2% solution) .........................................................1.0mL
FeSO 4 ·7H 2 O (2.5% solution)...................................................0.03mL
pH 7.3 ± 0.1 at 25°C
Reducing Agent:
Composition per 20.0mL:
Na 2 S·9H 2 O....................................................................................0.5g
Preparation of Reducing Agent: Add Na 2 S· 9H 2 O to distilled/de-
ionized water and bring volume to 20.0mL. Mix thoroughly. Gas with
100% N 2 for 20 min. Cap with a rubber stopper. Autoclave for 45 min
at 15 psi pressure–121°C. Use freshly prepared solution.
Yeast Extract Solution:
Composition per 10.0mL:
Yeast extract..................................................................................1.0g
Preparation of Yeast Extract Solution: Add yeast extract to dis-
tilled/deionized water and bring volume to 10.0mL. Mix thoroughly.
Autoclave for 45 min at 15 psi pressure–121°C. Cool to 25°C.
Trace Minerals:
Composition per liter:
Nitrilotriacetic acid.....................................................................12.8g
NaCl..............................................................................................1.0g
CoCl 2 ·6H 2 O................................................................................0.16g
CaCl 2 ·2H 2 O ..................................................................................0.1g
FeSO 4 ·7H 2 O..................................................................................0.1g
MnCl 2 ·4H 2 O .................................................................................0.1g
ZnCl 2 .............................................................................................0.1g
NiSO 4 ·6H 2 O..............................................................................0.026g
CuCl 2 ..........................................................................................0.02g
Na 2 SeO 3 ......................................................................................0.02g
H 3 BO 3 .........................................................................................0.01g
Na 2 MoO 4 ·2H 2 O..........................................................................0.01g
Preparation of Trace Minerals: 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.
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.
Preparation of Medium: Add components, except yeast extract so-
lution and reducing agent, to distilled/deionized water and bring vol-
ume to 1.0L. Mix thoroughly. Gently heat and bring to boiling. Cool
under 90% N 2 + 10% CO 2 . Anaerobically distribute into tubes in
6.0mL volumes. Autoclave for 45 min at 15 psi pressure–121°C. Asep-
tically add 0.06mL of sterile yeast extract solution to each tube. Mix
thoroughly. Immediately prior to inoculation, aseptically add 0.12mL
of sterile reducing agent to each tube. Mix thoroughly.
Use:  For the cultivation and maintenance of Clostridium thermosulfuro-
genes.