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培養基配方

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Creatinine/NMH Medium

[所屬分類:培養基配方] [發布時間:2021-11-15] [發布人:網站管理員2] [閱讀次數:] [返回]

山東拓(tuo)普生物(wu)工程有限公司(si) 培(pei)養基配方(fang) //ljxinglong.com

Composition per 1100.0mL:

Yeast extract........................................................5.0g

NaCl....................................................................1.0g

NaHCO3..............................................................1.0g

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

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

MnCl2·4H2O .....................................................0.06g

CaSO4·2H2O .....................................................0.05g

FeSO4·7H2O......................................................0.01g

Na2SeO3·5H2O................................................26.0μg

Vitamin B12 .....................................................20.0μg

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

Phosphate solution...................................... 100.0mL

Creatinine solution...................................... 100.0mL

Trace elements solution SL-4 ....................... 10.0mL

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

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

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

pH 7.0 ± 0.2 at 25°C

Phosphate Solution:

Composition per 100.0mL:

K2HPO4.............................................................5.33g

KH2PO4.............................................................2.64g

Preparation of Phosphate Solution: Add com

ponents to distilled/deionized water and bring vol

ume to 100.0mL. Mix thoroughly. Sparge with 100%

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

Creatinine Solution:

Composition per 100.0mL:

Creatinine............................................................5.5g

Preparation of Creatinine Solution: Add crea

tinine to distilled/deionized water and bring volume

to 100.0mL. Mix thoroughly. Sparge with 100% N2.

Filter sterilize.

Trace Elements Solution SL-4:

Composition per liter:

EDTA ..................................................................0.5g

FeSO4·7H2O....................................................... 0.2g

Trace elements solution SL-6 ..................... 100.0mL

Trace Elements Solution SL-6:

Composition per liter:

MnCl2·4H2O .......................................................0.5g

H3BO3 .................................................................0.3g

CoCl2·6H2O ........................................................0.2g

ZnSO4·7H2O ...................................................... 0.1g

Na2MoO4·2H2O ............................................... 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.

Preparation of Trace Elements Solution SL-4:

Add components to distilled/deionized water and

bring volume to 1.0L. Mix thoroughly.

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.

L-Cysteine·HCl Solution:

Composition per 10.0mL:

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

Preparation of L-Cysteine·HCl Solution: Add

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

volume to 10.0mL. Mix thoroughly. Sparge with

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

121°C.

Na2S·9H2O Solution:

Composition per 10.0mL:

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

Preparation of Na2S·9H2O Solution: Add

Na2S·9H2O to distilled/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: Add components, ex

cept creatinine solution, phosphate solution, L-cys

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

distilled/deionized water and bring volume to

880.0mL Mix thoroughly. Adjust pH to 7.0. Auto

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

ly and anaerobically add 100.0mL of sterile

phosphate solution, 10.0mL of sterile L-cysteine·HCl

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

Immediately prior to use, aseptically and anaerobi

cally add 100.0mL of sterile creatinine solution. Mix

thoroughly. Aseptically and anaerobically distribute

into tubes or bottles.

Use: For the cultivation and maintenance of Clostrid

ium species.

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Creatinine/NMH Medium