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Sludge Medium for Methanobacteria, pH 7.9

[所属分类:培养基配方] [发布时间:2021-8-17] [发布人:网站管理员2] [阅读次数:] [返回]

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

Composition per liter:

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

Sodium formate............................................................................. 2.0g

Sodium acetate.............................................................................. 1.0g

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

L-Cysteine·HCl·H2O ..................................................................... 0.5g

KH2PO4......................................................................................... 0.5g

Na2S·9H2O.................................................................................... 0.5g

MgSO4·7H2O ................................................................................ 0.4g

NaCl .............................................................................................. 0.4g

NH4Cl ........................................................................................... 0.4g

CaCl2·2H2O................................................................................. 0.05g

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

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

Sludge fluid..............................................................................50.0mL

Fatty acid mixture....................................................................20.0mL

Trace elements solution SL-6 ....................................................1.0mL

pH 7.9 ± 0.2 at 25°C

Sludge Fluid:

Composition per 100.0mL:

Yeast extract.................................................................................. 0.4g

Sludge ....................................................................................100.0mL

Preparation of Sludge Fluid: Add yeast extract to a concentration

of 0.4% to sludge taken from an anaerobic digester. Gas with 100% N2

for 5 min. Incubate at 37°C for 24 hr. Centrifuge at 13,000 × g. Remove

the supernatant fluid. Gas with 100% N2 for 5 min. Autoclave for 15

min at 15 psi pressure–121°C. Store at 25°C protected from light.

Fatty Acid Mixture:

Composition per 20.0mL:

α-Methylbutyric acid.................................................................... 0.5g

Isobutyric acid .............................................................................. 0.5g

Isovaleric acid............................................................................... 0.5g

Valeric acid ................................................................................... 0.5g

Preparation of Fatty Acid Mixture: Add components to distilled/

deionized water and bring volume to 20.0mL. Mix thoroughly. Adjust

pH to 7.5 with concentrated NaOH.

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. Ad

just pH to 3.4.

Preparation of Medium: Prepare and dispense medium under 80%

N2 + 20% CO2. Add components to distilled/deionized water and bring

volume to 1.0L. Adjust pH to 7.9. Distribute anaerobically into tubes

or bottles with aluminum seals. Autoclave for 15 min at 15 psi pres

sure–121°C with fast exhaust.

Use: For the cultivation of Methanobacterium alcaliphilum and Metha

nobacterium thermoalcalip.

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Sludge Medium for Methanobacteria, pH 7.9