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Xylan Medium

[所属分类:培养基配方] [发布时间:2021-10-15] [发布人:网站管理员2] [阅读次数:] [返回]

Xylan Medium

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

Composition per liter:

Xylan........................................................................................... 30.0g

Agar ............................................................................................ 12.0g

Peptone.......................................................................................... 2.0g

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

L-Cysteine·HCl·H2O ................................................................... 0.25g

Na2S·9H2O.................................................................................. 0.25g

Rumen fluid ...........................................................................400.0mL

NaHCO3 solution .....................................................................40.0mL

Mineral solution I.....................................................................25.0mL

Mineral solution II ...................................................................25.0mL

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

VFA solution............................................................................10.0mL

Hemin solution.........................................................................10.0mL

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

pH 7.0 ± 0.2 at 25°C

NaHCO3 Solution:

Composition per 100.0mL:

NaHCO3 ...................................................................................... 3.96g

Preparation of NaHCO3 Solution: Add NaHCO3 to distilled/de

ionized water and bring volume to 100.0mL. Mix thoroughly. Gas with

100% CO2.

Mineral Solution I:

Composition per liter:

K2HPO4......................................................................................... 3.0g

Preparation of Mineral Solution I: Add K2HPO4 to distilled/de

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

Mineral Solution II:

Composition per liter:

Sodium citrate ............................................................................. 20.0g

NaCl ............................................................................................ 12.0g

KH2PO4......................................................................................... 6.0g

MgCl2·6H2O.................................................................................. 2.0g

CaCl2............................................................................................. 1.2g

Preparation of Mineral Solution II: Add components to dis

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

Wolfe’s Vitamin Solution:

Composition per liter:

Pyridoxine·HCl ........................................................................10.0mg

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 .....................................................................100.0μg

Preparation of Wolfe’s Vitamin Solution: Add components to

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

VFA Solution:

Composition per liter:

Acetic acid .............................................................................178.3mL

Propionic acid ..........................................................................59.6mL

n-Butyric acid ..........................................................................38.4mL

Isobutyric acid ...........................................................................9.5mL

n-Valeric acid.............................................................................9.4mL

Isovaleric acid............................................................................9.3mL

DL-α-Methylbutyric acid ...........................................................4.4mL

Preparation of VFA Solution: Add components to distilled/deion

ized water and bring volume to approximately 500.0mL. Adjust pH to

7.5 with NaOH. Mix thoroughly. Bring volume to 1.0L with distilled/

deionized water.

Hemin Solution:

Composition per 100.0mL:

Hemin ......................................................................................... 0.01g

Preparation of Hemin Solution: Add hemin to 100.0mL of 0.01N

NaOH. Mix thoroughly.

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 Na2S·9H2O,

NaCHO3,and L-cysteine·HCl·H2O solutions, to distilled/deionized wa

ter and bring volume to 1.0L. Mix thoroughly. Gently heat and bring to

boiling. Cool under 80% N2 + 20% CO2. Add L-cysteine·HCl·H2O and

Na2S·9H2O. Add sufficient NaCHO3 solution to bring pH to 7.2 under

80% N2 + 20% CO2. Anaerobically distribute into tubes under 80% N2

+ 20% CO2. Autoclave for 15 min at 15 psi pressure–121°C.

Use: For the cultivation and maintenance of Clostridium xylanolyti

cum and other microorganisms that can utilize xylan as a carbon

source.

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Xylan Medium