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Sporomusa Medium

[所属分类:培养基配方] [发布时间:2021-8-23] [发布人:网站管理员2] [阅读次数:] [返回]

Sporomusa Medium

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

Composition per 877.0mL:

NaCl............................................................................................ 2.25g

Pancreatic digest of casein............................................................ 2.0g

Yeast extract.................................................................................. 2.0g

MgSO4·7H2O............................................................................... .0.5g

NH4Cl ........................................................................................... 0.5g

K2HPO4....................................................................................... 0.35g

KH2PO4....................................................................................... 0.23g

CaCl2·2H2O .............................................................................. 0.025g

FeSO4·7H2O............................................................................... 2.0mg

Resazurin ................................................................................... 1.0mg

NaHSeO3 ..................................................................................15.0μg

NaHCO3 solution.....................................................................50.0mL

Glycine betaine solution ..........................................................50.0mL

Wolfe’s vitamin solution..........................................................10.0mL

L-Cysteine·HCl·H2O solution ..................................................10.0mL

Trace elements solution SL-6 ....................................................3.0mL

pH 7.0–7.2 at 25°C

NaHCO3 Solution:

Composition per 50.0mL:

NaHCO3........................................................................................ 4.0g

Preparation of NaHCO3 Solution: Add the NaHCO3 to distilled/

deionized water and bring volume to 50.0mL. Mix thoroughly. Gas un

der 80% N2 + 20% CO2 for 20 min.

Glycine Betaine Solution:

Composition per 50.0mL:

Glycine betaine ............................................................................. 5.0g

Preparation of Glycine Betaine Solution: Add the glycine be

taine to distilled/deionized water and bring volume to 50.0mL. Mix

thoroughly. Filter sterilize. Aseptically gas under 80% N2 + 20% CO2.

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.

L-Cysteine·HCl·H2O Solution:

Composition per 10.0mL:

L-Cysteine·HCl·H2O ..................................................................... 0.3g

Preparation of L-Cysteine·HCl·H2O Solution: Add L-cyste

ine·HCl·H2O to distilled/deionized water and bring volume to 10.0mL.

Mix thoroughly. Filter sterilize. Aseptically gas under 100% N2.

Trace Elements Solution SL-6:

Composition per liter:

H3BO3 ........................................................................................... 0.3g

CoCl2·6H2O .................................................................................. 0.2g

ZnSO4·7H2O ................................................................................. 0.1g

MnCl2·4H2O................................................................................ 0.03g

Na2MoO4·H2O ............................................................................ 0.03g

NiCl2·6H2O ................................................................................. 0.02g

CuCl2·2H2O................................................................................. 0.01g

Preparation of Trace Elements Solution SL-6: Add components

to distilled/deionized water and bring volume to 1.0L. Mix thoroughly.

Adjust pH to 3.4.

Preparation of Medium: Add components—except NaHCO3 solu

tion, glycine betaine solution, and L-cysteine·HCl·H2O solution—to

distilled/deionized water and bring volume to 890.0mL. Mix thorough

ly. Gently heat and bring to boiling. Continue boiling for 5 min. Cool

rapidly to 25°C under 80% N2 + 20% CO2. Add 50.0mL of NaHCO3

solution. Mix thoroughly. Autoclave anaerobically for 15 min at 15 psi

pressure–121°C. Cool to 25°C. Aseptically add sterile glycine betaine

solution. Mix thoroughly. Immediately prior to inoculation, aseptically

and anaerobically add L-cysteine·HCl·H2O solution.

Use: For the cultivation and maintenance of Sporomusa ovata and

Sporomusa sphaeroides.

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Sporomusa Medium