Chopped Meat Medium with Menadione

[所属分类:培养基配方] [发布时间:2020-4-26] [发布人:] [阅读次数:] [返回]

山东拓普生物工程有限公司

Shandong Tuopu Biol-Engineering Co.,Ltd

Chopped Meat Medium with Menadione

培养基配方
Composition per 1230.0mL:
Peptone........................................................................30.0g
K2HPO4 .........................................................................5.0g
Yeast extract...................................................................5.0g
L- Cysteine·HCl·H2O...........................................................0.5g
Chopped meat extract filtrate...................................................1.0L
Chopped meat extract solids..................................................200.0mL
Resazurin (0.025% solution) ...................................................4.0mL
Menadione solution ...........................................................0.25mL
pH 7.0 ± 0.2 at 25°C
Chopped Meat Extract:
Composition per liter:
Beef or horse meat............................................................500.0g
NaOH (1N solution)............................................................25.0mL
Preparation of Chopped Meat Extract: Use lean beef or horse
meat. Remove fat and connective tissue. Grind. Add meat and NaOH
to distilled/deionized water and bring volume to 1.0L. Gently heat and
bring to boiling while stirring. Cool to 25°C. Remove fat from surface.
Filter. Reserve ground meat particles and filtrate. Add distilled/deion-
ized water to filtrate and bring volume to 1.0L.
Menadione Solution:
Composition per liter:
Menadione (vitamin K3 ).....................................................50.0μg
Ethanol (20% solution).......................................................25.0mL
Preparation of Menadione Solution: Dissolve menadione in eth-
anol. Filter sterilize.
Preparation of Medium: To 1.0L of chopped meat extract filtrate,
add the remaining components, except the  L- cysteine·HCl·H2O, me-
nadione solution, and chopped meat solids. Mix thoroughly. Gently
heat to boiling. Cool to room temperature. Add the  L-
cysteine·HCl·H2O. Adjust pH to 7.0. Distribute 1 part chopped meat
solids (by volume) and 5 parts of liquid (by volume) into tubes under
O2 -free 97% N2 + 3% H2 . Cap with rubber stoppers and place tubes
in a press. Autoclave for 15 min at 15 psi pressure–121°C with fast
exhaust. Prior to inoculation, add 0.25mL of menadione solution to
each tube containing approximately 5.0mL of chopped meat medium.
Use:  For the cultivation and maintenance of Bacteroides gingivalis,
Bacteroides macacae, and Porphyromonas gingivalis.