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Vulcanithermus Medium (DSMZ Medium 977)

[所属分类:培养基配方] [发布时间:2021-10-12] [发布人:网站管理员2] [阅读次数:] [返回]

Vulcanithermus Medium

(DSMZ Medium 977)

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

Composition per liter:

NaCl ........................................................................................... 25.0g

NH4Cl ........................................................................................ 0.33g

KCl ............................................................................................. 0.33g

Calcium chloride solution........................................................10.0mL

Magnesium chloride solution...................................................10.0mL

Potassium nitrate solution........................................................10.0mL

Tryptone solution .....................................................................10.0mL

Sucrose solution.......................................................................10.0mL

Yeast extract solution...............................................................10.0mL

PIPES solution .......................................................................... 3.6mL

Vitamin solution.........................................................................1.0mL

Trace elements solution .............................................................1.0mL

pH 6.8 ± 0.2 at 25°C

Trace Elements Solution:

Composition per liter:

MgSO4·7H2O ................................................................................ 3.0g

Nitrilotriacetic acid ....................................................................... 1.5g

NaCl .............................................................................................. 1.0g

MnSO4·2H2O ................................................................................ 0.5g

CoSO4·7H2O ............................................................................... 0.18g

ZnSO4·7H2O ............................................................................... 0.18g

CaCl2·2H2O................................................................................... 0.1g

FeSO4·7H2O.................................................................................. 0.1g

NiCl2·6H2O ............................................................................... 0.025g

KAl(SO4)2·12H2O....................................................................... 0.02g

H3BO3 ......................................................................................... 0.01g

Na2MoO4·4H2O .......................................................................... 0.01g

CuSO4·5H2O ............................................................................... 0.01g

Na2SeO3·5H2O...........................................................................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 remaining components. Add distilled/deionized

water to 1.0L. Mix thoroughly.

Vitamin Solution:

Composition per liter:

Pyridoxine-HCl........................................................................10.0mg

Thiamine-HCl·2H2O..................................................................5.0mg

Riboflavin ..................................................................................5.0mg

Nicotinic acid.............................................................................5.0mg

D-Ca-pantothenate......................................................................5.0mg

p-Aminobenzoic acid.................................................................5.0mg

Lipoic acid .................................................................................5.0mg

Biotin .........................................................................................2.0mg

Folic acid ................................................................................... 2.0mg

Vitamin B12 ................................................................................ 0.1mg

Preparation of Vitamin Solution: Add components to distilled/

deionized water and bring volume to 1.0L. Mix thoroughly. Sparge

with 80% H2 + 20% CO2. Filter sterilize.

PIPES Solution:

Composition per liter:

PIPES (Piperazine-N,N'-bis[2-ethane-sulfonic acid]) ................. 3.62

Preparation of PIPES Solution: Add PIPES to distilled/deionized

water and bring volume to 1.0L. Mix thoroughly.

Yeast Extract Solution:

Composition per 10.0mL:

Yeast extract ................................................................................. 0.5g

Preparation of Yeast Extract Solution: Add yeast extract to dis

tilled/deionized water and bring volume to 10.0mL. Mix thoroughly.

Autoclave for 15 min at 15 psi pressure–121°C. Cool to room temper

ature. Sparge with 100% N2.

Sucrose Solution:

Composition per 10.0mL:

Sucrose ......................................................................................... 1.0g

Preparation of Sucrose Solution: Add sucrose to distilled/deion

ized water and bring volume to 10.0mL. Mix thoroughly. Autoclave for

15 min at 15 psi pressure–121°C. Cool to room temperature. Sparge

with 100% N2.

Tryptone Solution:

Composition per 10.0mL:

Tryptone....................................................................................... 1.0g

Preparation of Tryptone Solution: Add tryptone to distilled/de

ionized water and bring volume to 10.0mL. Mix thoroughly. Autoclave

for 15 min at 15 psi pressure–121°C. Cool to room temperature. Sparge

with 100% N2.

Postassium Nitrate Solution:

Composition per 10.0mL:

KNO3 .......................................................................................... 0.33g

Preparation of Potassium Nitrate Solution: Add KNO3 to dis

tilled/deionized water and bring volume to 10.0mL. Mix thoroughly.

Autoclave for 15 min at 15 psi pressure–121°C. Cool to room temper

ature. Sparge with 100% N2.

Magnesium Chloride Solution:

Composition per 10.0mL:

MgCl2·6H2O ............................................................................... 0.33g

Preparation of Magnesium Chloride Solution: Add0.33g of

MgCl2·6H2O to distilled/deionized water and bring volume to 10.0mL.

Mix thoroughly. Autoclave for 15 min at 15 psi pressure–121°C. Cool

to room temperature. Sparge with 100% N2.

Calcium Chloride Solution:

Composition per 10.0mL:

CaCl2·2H2O ................................................................................ 0.33g

Preparation of Calcium Chloride Solution: Add CaCl2·2H2O

to distilled/deionized water and bring volume to 10.0mL. Mix thor

oughly. Autoclave for 15 min at 15 psi pressure–121°C. Cool to room

temperature. Sparge with 100% N2.

Preparation of Medium: Add components, except calcium chlo

ride, magnesium chloride, potassium nitrate, tryptone, yeast extract, vi

tamin, and sucrose solutions, to distilled/deionized water and bring

volume to 940.0mL. Mix thoroughly. Sparge with 100% N2. Adjust pH

to 6.8. Dispense into vessels suitable for anaerobic growth (Hungate

tubes or serum bottles) under an atmosphere of 100% N2. Autoclave

for 15 min at 15 psi pressure–121°C. Cool to room temperature. Add

calcium chloride, magnesium chloride, potassium nitrate, tryptone,

yeast extract, vitamin, and sucrose solutions. Mix thoroughly. Adjust

pH to 6.8. Aseptically dispense into culture vessels.

Use: For the cultivation of Vulcanithermus mediatlanticus.

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Vulcanithermus Medium (DSMZ Medium 977)