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M10 Broth

[所属分类:培养基配方] [发布时间:2021-11-29] [发布人:网站管理员2] [阅读次数:] [返回]

M10 Broth

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

Composition per liter:

Pancreatic digest of casein..................................2.0g

Yeast extract........................................................2.0g

Cellobiose ...........................................................1.0g

Glucose ...............................................................1.0g

Maltose................................................................1.0g

Starch ..................................................................1.0g

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

Mineral solution I........................................ 100.0mL

Mineral solution II ...................................... 100.0mL

Na2CO3 solution ........................................... 50.0mL

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

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

Volatile fatty acid mixture............................... 3.1mL

pH 6.8 ± 0.2 at 25°C

Mineral Solution I:

Composition per 100.0mL:

K2HPO4...............................................................0.2g

Preparation of Mineral Solution I: Add

K2HPO4 to distilled/deionized water and bring vol

ume to 100.0mL. Mix thoroughly.

Mineral Solution II:

Composition per 100.0mL:

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

(NH4)2SO4...........................................................0.4g

KH2PO4...............................................................0.3g

MgSO4·7H2O ....................................................0.09g

CaCl2.................................................................0.05g

Preparation of Mineral Solution II: Add com

ponents to distilled/deionized water and bring vol

ume to 100.0mL. Mix thoroughly.

Na2CO3 Solution:

Composition per 100.0mL:

Na2CO3 ...............................................................8.0g

Preparation of Na2CO3 Solution: Add Na2CO3

to distilled/deionized water and bring volume to

100.0mL. Mix thoroughly. Sparge with 100% CO2.

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

Hemin Solution:

Composition per 100.0mL:

Hemin .................................................................1.0g

NaOH (1N solution)...................................... 10.0mL

Preparation of Hemin Solution: Add compo

nents to 100.0mL of distilled/deionized water. Mix

thoroughly.

L-Cysteine·HCl Solution:

Composition per 10.0mL:

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

Preparation of L-Cysteine·HCl Solution: Add

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

volume to 10.0mL. Mix thoroughly. Autoclave under

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

Volatile Fatty Acid Mixture:

Composition per 31.0mL:

Acetic acid .................................................... 17.0mL

Propionic acid ................................................. 6.0mL

Butyric acid..................................................... 4.0mL

DL-α-Methylbutyric acid ................................ 1.0mL

Isobutyric acid ................................................ 1.0mL

Isovaleric acid................................................. 1.0mL

n-Valeric acid.................................................. 1.0mL

Preparation of Volatile Fatty Acid Mixture:

Combine components. Mix thoroughly. Store under

100% N2.

Preparation of Medium: Prepare and dispense

medium under 100% CO2. Add components, except

L-cysteine·HCl solution and Na2CO3 solution, to dis

tilled/deionized water and bring volume to 930.0mL.

Mix thoroughly. Sparge with 100% CO2. Adjust pH

to 6.8 with 1N NaOH. Distribute anaerobically

9.3mL volumes into Hungate tubes. Autoclave for 15

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

obically add 0.2mL of sterile L-cysteine·HCl solution

and 0.5mL of sterile Na2CO3 solution. Check that fi

nal pH is 6.8. (Note: if not properly gassed with

100% CO2, the medium pH can be as high as 9.5.)

Use: For the cultivation of Enterococcus species,

Lactobacillus species and Streptococcus species.

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M10 Broth