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WMC Medium

[所属分类:培养基配方] [发布时间:2021-10-14] [发布人:网站管理员2] [阅读次数:] [返回]

WMC Medium

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

Composition per 1010.0mL:

NaHCO3........................................................................................ 5.0g

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

L-Cysteine ..................................................................................... 0.5g

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

Mineral salt solution 2xW......................................................500.0mL

LIP solution..............................................................................50.0mL

TYC solution............................................................................50.0mL

Trace elements solution ...........................................................10.0mL

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

pH 7.2 ± 0.2 at 25°C

Mineral Solution 2xW:

Composition per liter:

NaCl ............................................................................................ 40.0g

MgCl2·6H2O.................................................................................. 5.6g

MgSO4·7H2O ................................................................................ 0.7g

KCl.............................................................................................. 0.68g

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

CaCl2·2H2O................................................................................. 0.28g

K2HPO4...................................................................................... .0.28g

Preparation of Mineral Solution 2xW: Add components to dis

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

sterilize. Store at room temperature in the dark.

LIP Solution:

Composition per liter:

L-Isoleucine................................................................................. 10.0g

L-Leucine ...................................................................................... 5.0g

Pantothenate.................................................................................. 0.1g

Preparation of LIP Solution: Add components to distilled/deion

ized water and bring volume to 1.0L. Mix thoroughly. May be stored

unsterilized at −20°C.

TYC Solution:

Composition per liter:

Casamino acids ......................................................................... 100.0g

Yeast extract................................................................................ 50.0g

L-Tryptophan................................................................................. 1.0g

Preparation of TYC Solution: Add components to distilled/de

ionized water and bring volume to 1.0L. Mix thoroughly. Sparge with

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

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

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

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

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

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

Na2MoO4·2H2O ......................................................................... 0.01g

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

Na2SeO3·5H2O...........................................................................0.3mg

Preparation of Trace Elements Solution: Add nitrilotriacetic

acid to 500.0mL of distilled/deionized water. Adjust pH to 6.5 with

KOH. Add remaining components. Add distilled/deionized water to

1.0L.

Na2S·9H2O Solution:

Composition per 10.0mL:

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

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

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

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

Preparation of Medium: Prepare and dispense medium under 80%

H2 + 20% CO2. Add components, except TYC solution and Na2S·9H2O

solution, to distilled/deionized water and bring volume to 940.0mL.

Mix thoroughly. Sparge with 80% H2 + 20% CO2. Autoclave for 15

min at 15 psi pressure–121°C. Aseptically and anaerobically add

50.0mL of sterile TYC solution and 10.0mL of sterile Na2S·9H2O so

lution. Mix thoroughly.

Use: For the cultivation and maintenance of Methanococcus voltae.

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WMC Medium