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培養基配方

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PY Medium with Glucose (Peptone Yeast Extract Medium with Glucose) (PYG Medium)

[所屬分類:培養基配方] [發布時間:2021-12-13] [發布人:網站管理員2] [閱讀次數:] [返回]

PY Medium with Glucose

(Peptone Yeast Extract

Medium with Glucose)

(PYG Medium)

山東拓普生物工程有限公司培養基配方 //ljxinglong.com

Composition per liter:

Glucose .............................................................10.0g

Yeast extract......................................................10.0g

Peptone................................................................5.0g

Pancreatic digest of casein..................................5.0g

L-cysteine·HCl·H2O ............................................0.5g

Salts solution................................................. 40.0mL

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

Resazurin solution .......................................... 4.0mL

Vitamin K1 solution ........................................ 0.2mL

pH 6.9 ± 0.2 at 25°C

Salts Solution:

Composition per liter:

NaHCO3............................................................10.0g

NaCl....................................................................2.0g

K2HPO4...............................................................1.0g

KH2PO4...............................................................1.0g

CaCl2, anhydrous................................................0.2g

MgSO4 ................................................................0.2g

Preparation of Salts Solution: Add CaCl2 and

MgSO4 to 300.0mL of distilled/deionized water. Mix

thoroughly until dissolved. Bring volume to

800.0mL with distilled/deionized water. Add remain

ing components while stirring. Bring volume to 1.0L.

Mix thoroughly. Store at 4°C.

Hemin Solution:

Composition per 100.0mL:

Hemin ...............................................................0.05g

NaOH (1N solution)........................................ 1.0mL

Preparation of Hemin Solution: Add hemin to

NaOH solution. Mix thoroughly. Adjust volume to

100.0mL with distilled/deionized water. Autoclave

for 15 min at 15 psi pressure–121°C. Cool to 45°–

50°C.

Resazurin Solution:

Composition per 44.0mL:

Resazurin ........................................................0.044g

Preparation of Resazurin Solution: Add resa

zurin to distilled/deionized water and bring volume

to 44.0mL. Mix thoroughly.

Vitamin K1 Solution:

Composition per 30.0mL:

Vitamin K1 ........................................................0.15g

Ethanol (95% solution)................................. 30.0mL

Preparation of Vitamin K1 Solution: Add vita

min K1 to ethanol. Mix thoroughly. Store in brown

bottle and keep under refrigeration. Discard after 1

month.

Preparation of Medium: Add components—ex

cept L-cysteine·HCl·H2O, hemin solution, and vita

min K1 solution—to distilled/deionized water and

bring volume to 989.8mL. Mix thoroughly. Gently

heat and bring to boiling under 80% N2 + 10% CO2

+ 10% H2. Continue boiling until resazurin turns col

orless, indicating reduction. Cool to 45°–50°C. Add

the L-cysteine·HCl·H2O, hemin solution, and vitamin

K1 solution. Adjust pH to 6.9 if necessary. Anaerobi

cally distribute into tubes under 80% N2 + 10% CO2

+ 10% H2. Cap the tubes with rubber stoppers. Place

tubes in a press. Autoclave for 15 min at 15 psi pres

sure–121°C with fast exhaust.

Use: For the cultivation and maintenance of

Clostridium species.

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PY Medium with Glucose (Peptone Yeast Extract Medium with Glucose) (PYG Medium)