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GV Medium

[所属分类:培养基配方] [发布时间:2021-11-22] [发布人:网站管理员2] [阅读次数:] [返回]

GV Medium

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

Composition per 1061.0mL:

NaHCO3 ..............................................................3.0g

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

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

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

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

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

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

Resazurin .........................................................0.5mg

Substrate solution.......................................... 20.0mL

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

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

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

MgSO4·7H2O solution .................................. 10.0mL

CaCl2·2H2O solution..................................... 10.0mL

Selenite/tungstate solution .............................. 1.0mL

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

pH 7.0 ± 0.2 at 25°C

Substrate Solution:

Composition per 20.0mL:

Sodium citrate.....................................................6.0g

Preparation of Substrate Solution: Add sodi

um citrate to distilled/deionized water and bring vol

ume to 20.0mL. Mix thoroughly. Sparge with 80%

N2 + 20% CO2. Autoclave for 15 min at 15 psi pres

sure–121°C.

Vitamin Solution:

Composition per liter:

Pyridoxine-HCl..............................................10.0mg

Calcium DL-pantothenate.................................5.0mg

Lipoic acid .......................................................5.0mg

Nicotinic acid...................................................5.0mg

p-Aminobenzoic acid.......................................5.0mg

Riboflavin ........................................................5.0mg

Thiamine-HCl ..................................................5.0mg

Biotin ...............................................................2.0mg

Folic acid..........................................................2.0mg

Vitamin B12 ......................................................0.1mg

Preparation of Vitamin Solution: Add compo

nents to distilled/deionized water and bring volume

to 1.0L. Mix thoroughly. Sparge with 80% N2 + 20%

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

L-Cysteine·HCl Solution:

Composition per 10.0mL:

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

Preparation of L-Cysteine·HCl Solution: Add

L-cysteine·HCl to distilled/deionized water and bring

volume to 10.0mL. Mix thoroughly. Sparge with

80% N2 + 20% CO2. 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 distilled/deionized water and bring

volume to 10.0mL. Mix thoroughly. Gas under 80%

N2 + 20% CO2. Autoclave for 15 min at 15 psi pres

sure–121°C.

MgSO4·7H2O Solution:

Composition per 10.0mL:

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

Preparation of MgSO4·7H2O Solution: Add

MgSO4·7H2O to distilled/deionized water and bring

volume to 10.0mL. Mix thoroughly. Sparge with

80% N2 + 20% CO2. Autoclave for 15 min at 15 psi

pressure–121°C.

CaCl2·2H2O Solution:

Composition per 10.0mL:

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

Preparation of CaCl2·2H2O Solution: Add

CaCl2·2H2O to distilled/deionized water and bring

volume to 10.0mL. Mix thoroughly. Sparge with

80% N2 + 20% CO2. Autoclave for 15 min at 15 psi

pressure–121°C.

Selenite-Tungstate Solution:

Composition per liter:

NaOH..................................................................0.5g

Na2WO4·2H2O ...................................................4mg

Na2SeO3·5H2O ...................................................3mg

Preparation of Selenite-Tungstate Solution:

Add components to distilled/deionized water and

bring volume to 1.0L. Mix thoroughly. Sparge with

80% N2 + 20% CO2. Autoclave for 15 min at 15 psi

pressure–121°C.

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

thoroughly. Gas under 80% N2 + 20% CO2. Auto

clave for 15 min at 15 psi pressure–121°C.

Preparation of Medium: Prepare and dispense

medium under 80% N2 + 20% CO2. Add components,

except substrate solution, MgSO4·7H2O solution,

CaCl2·2H2O solution, selenite/tungstate solution, L-cys

teine·HCl solution, and Na2S·9H2O solution, to dis

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

thoroughly. Sparge with 80% N2 + 20% CO2. Auto

clave for 15 min at 15 psi pressure–121°C. Aseptically

and anaerobically add 20.0mL of sterile substrate solu

tion, 10.0mL of sterile MgSO4·7H2O solution, 10.0mL

of sterile CaCl2·2H2O solution, 1.0mL of sterile selen

ite/tungstate solution, 10.0mL of sterile L-cysteine·HCl

solution, and 10.0mL of sterile Na2S·9H2O solution.

Mix thoroughly. Adjust pH to 7.0 to 7.2. Aseptically

and anaerobically distribute into sterile tubes or flasks.

Use: For the cultivation and maintenance of Campy

lobacter species.

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GV Medium