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Sporomusa silvacetica Medium (DSMZ Medium 777)

[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 silvacetica Medium

(DSMZ Medium 777)

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 ............................................................................................ 2.25g

Yeast extract.................................................................................. 1.0g

Casitone ........................................................................................ 1.0g

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

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

K2HPO4..................................................................................... 0.348g

KH2PO4..................................................................................... 0.227g

CaCl2·2H2O................................................................................. 0.25g

FeSO4·7H2O.............................................................................. 0.002g

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

NaHSeO3...................................................................................15.1μg

Cysteine solution......................................................................10.0mL

NaHCO3 solution .....................................................................10.0mL

Na2S·9H2O solution .................................................................10.0mL

Fructose solution......................................................................10.0mL

Vitamin solution.......................................................................10.0mL

Trace elements solution SL-10 ..................................................1.0mL

pH 6.6 ± 0.2 at 25°C

Cysteine Solution:

Composition per 10.0mL:

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

Preparation of Cysteine Solution: Add L-cysteine·HCl·H2O to

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

ly. Sparge with 100% N2. Autoclave for 15 min at 15 psi pressure–

121°C.

Na2S·9H2O Solution:

Composition per 10.0mL:

Na2S·9H2O.................................................................................... 0.3g

Preparation of Na2S·9H2O Solution: Add Na2S·9H2O to dis

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

Autoclave under 100% N2 for 15 min at 15 psi pressure–121°C. Cool

to room temperature.

NaHCO3 Solution:

Composition per 10.0mL:

NaHCO3 ........................................................................................ 1.5g

Preparation of NaHCO3 Solution: Add NaHCO3 to distilled/de

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

with 80% N2 + 20% CO2. Filter sterilize.

Fructose Solution:

Composition per 10.0mL:

Fructose......................................................................................... 5.0g

Preparation of Fructose Solution: Add fructose to distilled/de

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

with 100% N2. Autoclave for 15 min at 15 psi pressure–121°C. Cool to

room temperature.

Trace Elements Solution SL-10:

Composition per liter:

FeCl2·4H2O................................................................................... 1.5g

CoCl2·6H2O ........................................................................... 190.0mg

MnCl2·4H2O .......................................................................... 100.0mg

ZnCl2........................................................................................ 70.0mg

Na2MoO4·2H2O ....................................................................... 36.0mg

NiCl2·6H2O.............................................................................. 24.0mg

H3BO3 ........................................................................................ 6.0mg

CuCl2·2H2O ............................................................................... 2.0mg

HCl (25% solution)..................................................................10.0mL

Preparation of Trace Elements Solution SL-10: Add FeCl2·4H2O

to 10.0mL of HCl solution. Mix thoroughly. Add distilled/deionized

water and bring volume to 1.0L. Add remaining components. Mix thor

oughly. Sparge with 80% N2 + 20% CO2. Autoclave for 15 min at 15

psi pressure–121°C.

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.

Preparation of Medium: Prepare and dispense medium under 80%

N2 + 20% CO2 gas atmosphere. Add components, except NaHCO3 solu

tion, fructose solution, cysteine solution, vitamin solution, and

Na2S·9H2O solution, to distilled/deionized water and bring volume to

950.0mL. Mix thoroughly. Adjust pH to 6.5–6.7. Autoclave for 15 min

at 15 psi pressure–121°C. Cool to room temperature. Aseptically and an

aerobically add 10.0mL NaHCO3 solution, 10.0mL fructose solution,

10.0mL cysteine solution, 10.0mL vitamin solution, and 10.0mL

Na2S·9H2O solution. Mix thoroughly. Aseptically and anaerobically dis

tribute into sterile tubes or bottles.

Use: For the cultivation of Sporomusa silvacetica, Thermicanus aegyp

tius, Moorella thermoacetica, and Moorella mulderi.

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Sporomusa silvacetica Medium (DSMZ Medium 777)