PY Carbohydrate 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 Carbohydrate Medium

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

Composition per 110.0mL:

Polypeptone™...............................................................................1.0g
Yeast extract..................................................................................1.0g
Glucose .........................................................................................0.4g
Maltose..........................................................................................0.4g
Ribose ...........................................................................................0.4g
Starch, soluble...............................................................................0.4g
L -Cysteine·HCl·H 2 O ...................................................................0.09g
Resazurin .................................................................................0.16mg
Serum VFAH supplement........................................................10.0mL
NaHCO 3 solution.....................................................................10.0mL
Salts solution..............................................................................4.0mL
pH 7.2–7.5 at 25°C
Serum VFAH Supplement:
Composition per 107.5mL:
Rabbit serum..........................................................................100.0mL
Heme solution............................................................................5.0mL
VFA solution..............................................................................1.5mL
Thiamine pyrophosphate solution..............................................1.0mL
Preparation of Serum VFAH: Combine the four solutions. Mix
thoroughly. Filter sterilize.
Thiamine Pyrophosphate Solution:
Composition per 10.0mL:
Thiamine pyrophosphate.............................................................0.05g
Preparation of Thiamine Pyrophosphate Solution: Add thia-
mine pyrophosphate to distilled/deionized water and bring volume to
10.0mL. Mix thoroughly. Filter sterilize.
Heme Solution:
Composition per 100.0mL:
Hemin............................................................................................0.5g
NaOH (1N solution)...................................................................1.0mL
Preparation of Heme Solution: Add hemin to NaOH solution.
Mix thoroughly. Bring volume to 100.0mL with distilled/deionized
water.
VFA Solution:
Composition per 100.0mL:
Acetic acid, glacial.....................................................................5.0mL
n-Butyric acid ............................................................................4.0mL
n-Valeric acid.............................................................................1.0mL
Isobutyric acid............................................................................1.0mL
Isovaleric acid............................................................................1.0mL
n-Butyric acid ............................................................................1.0mL
Preparation of VFA Solution: Add components to distilled/deion-
ized water and bring volume to 100.0mL. Mix thoroughly. Adjust pH
to 7.0.
NaHCO 3 Solution:
Composition per 10.0mL:
NaHCO 3 ........................................................................................0.5g
Preparation of NaHCO 3 Solution: Add the NaHCO 3 to distilled/
deionized water and bring volume to 10.0mL. Mix thoroughly. Filter
sterilize.
Salts Solution:
Composition per 400.0mL:
K 2 HPO 4 .........................................................................................0.9g
NaCl..............................................................................................0.8g
KH 2 PO 4 .........................................................................................0.4g
MnCl 2 ·4H 2 O ...............................................................................0.16g
MgSO 4 ........................................................................................0.08g
Preparation of Salts Solution: Add components to distilled/deion-
ized water and bring volume to 400.0mL. Mix thoroughly.
Preparation of Medium: Add components, except serum VFAH
supplement and NaHCO 3 solution, to distilled/deionized water and
bring volume to 90.0mL. Mix thoroughly. Gently heat and bring to
boiling. Continue boiling until resazurin turns colorless, indicating re-
duction. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C.
Aseptically and anaerobically add 10.0mL of sterile serum VFAH sup-
plement and 10.0mL of sterile NaHCO 3 solution. Anaerobically dis-
tribute into sterile tubes or flasks under 100% N 2 .
Use: For the cultivation of oral treponemes.