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Peptone Yeast Extract Medium
(PY Medium)
(ATCC Medium 1524)

山東拓普生物工程有限公司 培養(yǎng)基配方 http://www.jlkcpj.cn

Composition per 950.0mL:

Yeast extract................................................................................10.0g
Peptone .........................................................................................5.0g
Pancreatic digest of casein............................................................5.0g
L -Cysteine·HCl·H 2 O.....................................................................0.5g
Salt solution.............................................................................40.0mL
Hemin solution.........................................................................10.0mL
Resazurin (0.025% solution) .....................................................4.0mL
Vitamin K 1 solution...................................................................0.2mL
pH 7.0 ± 0.2 at 25°C
Salt 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 Salt 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 and
bring volume to 100.0mL with distilled/deionized water. Mix thoroughly.
Autoclave for 15 min at 15 psi pressure–121°C. Cool to 45°–50°C.
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. Filter sterilize.
Preparation of Medium: Add components—except vitamin K 1 so-
lution, hemin solution, and  L -cysteine·HCl·H 2 O, to distilled/deionized
water and bring volume to 939.8mL. Gently heat and bring to boiling
under 80% N 2 + 10% H 2 + 10% CO 2 . Continue boiling until resazurin
turns colorless, indicating reduction. Cool to 45°–50°C. Add vitamin
K 1 solution, hemin solution, and  L -cysteine·HCl·H 2 O. Adjust pH to
7.0. Distribute into tubes under 80% N 2 + 10% H 2 + 10% CO 2 . Cap
with rubber stoppers. Place tubes in a press. Autoclave for 15 min at 15
psi pressure–121°C with fast exhaust.
Use:  For the cultivation and maintenance of Megasphaera cerevisiae
and Clostridium species.