Rumen Fluid Cellobiose Agar (RFC Agar)

[所属分类:培养基配方] [发布时间:2021-8-6] [发布人:网站管理员2] [阅读次数:] [返回]

Rumen Fluid Cellobiose Agar
(RFC Agar)

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

Composition per 10.0mL:

Rumen fluid cellobiose base medium........................................8.9mL
NaHCO 3 -rifampin solution........................................................1.0mL
Cellobiose solution ....................................................................0.1mL
Rumen Fluid Cellobiose Base Medium:
Composition per 89.0mL:
Noble agar.....................................................................................0.7g
Cysteine·HCl·H 2 O.........................................................................0.1g
Clarified rumen fluid ...............................................................30.0mL
Salts solution A........................................................................20.0mL
Salts solution B........................................................................20.0mL
Resazurin (0.1% solution)..........................................................0.1mL
pH 6.7–7.0 at 25°C
Preparation of Rumen Fluid Cellobiose Base Medium:  Add
components to distilled/deionized water and bring volume to 89.0mL.
Mix thoroughly. Gently heat and bring to boiling. Continue boiling until
resazurin turns colorless, indicating reduction. Anaerobically distribute
into tubes in 8.9mL volumes under 100% CO 2 . Cap tubes with rubber
stoppers. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C.
Salts Solution A:
Composition per liter:
CaCl 2 ...........................................................................................0.45g
MgSO 4 ........................................................................................0.45g
Preparation of Salts Solution A: Add components to distilled/de-
ionized water and bring volume to 1.0L. Mix thoroughly.
Salts Solution B:
Composition per liter:
NaCl..............................................................................................4.5g
(NH 4 ) 2 SO 4 .....................................................................................4.5g
KH 2 PO 4 .......................................................................................2.25g
K 2 HPO 4 .......................................................................................2.25g
Preparation of Salts Solution B: Add components to distilled/de-
ionized water and bring volume to 1.0L. Mix thoroughly.
NaHCO 3 -Rifampin Solution:
Composition per 10.0mL:
NaHCO 3 ........................................................................................0.5g
Rifampin ....................................................................................0.1mg
Preparation of NaHCO 3 -Rifampin Solution: Add components
to distilled/deionized water and bring volume to 10.0mL. Mix thor-
oughly. Filter sterilize.
Cellobiose Solution:
Composition per 10.0mL:
Cellobiose.....................................................................................1.0g
Preparation of Cellobiose Solution: Add cellobiose to distilled/
deionized water and bring volume to 10.0mL. Mix thoroughly. Filter
sterilize.
Preparation of Medium: To each tube containing 8.9mL of sterile
rumen fluid cellobiose base medium, aseptically add 1.0mL of sterile
NaHCO 3 -rifampin solution and 0.1mL of sterile cellobiose solution.
Mix thoroughly.
Use: For the selective isolation of rumen treponemes.