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

[所属分类:培养基配方] [发布时间:2021-7-16] [发布人:网站管理员2] [阅读次数:] [返回]
PY Medium with Glucose
(Peptone Yeast Extract Medium with Glucose)
(PYG 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:

Glucose .......................................................................................10.0g
Yeast extract................................................................................10.0g
Peptone..........................................................................................5.0g
Pancreatic digest of casein............................................................5.0g
L -cysteine·HCl·H 2 O......................................................................0.5g
Salts solution............................................................................40.0mL
Hemin solution.........................................................................10.0mL
Resazurin solution......................................................................4.0mL
Vitamin K 1 solution ...................................................................0.2mL
pH 6.9 ± 0.2 at 25°C
Salts Solution:
Composition per liter:
NaHCO 3 ......................................................................................10.0g
NaCl..............................................................................................2.0g
K 2 HPO 4 .........................................................................................1.0g
KH 2 PO 4 .........................................................................................1.0g
CaCl 2 , anhydrous..........................................................................0.2g
MgSO 4 ..........................................................................................0.2g
Preparation of Salts Solution: Add CaCl 2 and MgSO 4 to 300.0mL
of distilled/deionized water. Mix thoroughly until dissolved. Bring vol-
ume to 800.0mL with distilled/deionized water. Add remaining com-
ponents 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 resazurin to distilled/
deionized water and bring volume to 44.0mL. Mix thoroughly.
Vitamin K 1 Solution:
Composition per 30.0mL:
Vitamin K 1 ..................................................................................0.15g
Ethanol (95% solution)............................................................30.0mL
Preparation of Vitamin K 1 Solution:  Add vitamin K 1 to ethanol.
Mix thoroughly. Store in a brown bottle and keep under refrigeration.
Discard after 1 month.
Preparation of Medium: Add components—except hemin solu-
tion,  L -cysteine·HCl·H 2 O, and vitamin K 1 solution—to distilled/deion-
ized water and bring volume to 989.8mL. Mix thoroughly. Gently heat
and bring to boiling under 80% N 2 + 10% CO 2 + 10% H 2 . Continue
boiling until resazurin turns colorless, indicating reduction. Cool to
45°–50°C. Add the  L -cysteine·HCl·H 2 O, hemin solution, and vitamin
K 1 solution. Adjust pH to 6.9 if necessary. Anaerobically distribute into
tubes under 80% N 2 + 10% CO 2 + 10% H 2 . 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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