Chopped Meat Medium, Modified (DSMZ Medium 797)

[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, Modified
(DSMZ Medium 797)

培养基配方
Composition per 1230.0mL:
Chopped meat extract solids...................................................200.0g
Trypticase™....................................................................30.0g
Agar...........................................................................20.0g
K2HPO4 .........................................................................5.0g
Yeast extract...................................................................5.0g
L- Cysteine·HCl·H2O...........................................................0.5g
Chopped meat extract filtrate...................................................1.0L
Hemin solution................................................................10.0mL
Resazurin (0.025% solution) ...................................................4.0mL
Vitamin K 1 solution...........................................................0.2mL
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 both ground meat particles and filtrate. Add distilled/
deionized water to filtrate and bring volume to 1.0L.
Hemin Solution:
Composition per 100.0mL:
Hemin............................................................................0.05g
NaOH (1N solution)...............................................................1.0mL
Preparation of Hemin Solution: Add components to distilled/de-
ionized water and bring volume to 100.0mL. Mix thoroughly.
Vitamin K 1 Solution:
Composition per 30.0mL:
Ethanol (95% solution)............................................................30.0mL
Vitamin K 1 ......................................................................0.15mL
Preparation of Vitamin K 1 Solution: Mix components. Store so-
lution protected from light at 5°C. Discard after 1 month.
Preparation of Medium: To 1.0L of chopped meat extract filtrate,
add the remaining components, except the  L -cysteine·HCl·H2O, hemin
solution, vitamin K 1 solution, and chopped meat solids. Mix thor-
oughly. Gently heat to boiling. Boil for 5 min. Cool to 25°C while
sparging with 80% N2 + 10% H2  + 10% CO2 . Add the  L -
cysteine·HCl·H2O, hemin solution, and vitamin K 1 solution. Adjust pH
to 7.0. Distribute 1 part chopped meat solids (by volume) and 5 parts
of liquid (by volume) into tubes under 80% N2 + 10% H2  + 10% CO2 .
Autoclave for 15 min at 15 psi pressure–121°C.
Use:  For the cultivation of Dialister pneumosintes.