Chopped Meat Medium, Modified with Arginine

[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 with Arginine

培养基配方
Composition per 1230.0mL:
Pancreatic digest of casein..................................................30.0g
Peptone .....................................................................30.0g
Agar.........................................................................20.0g
Arginine.....................................................................5.0g
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
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
Tween™ 80..................................................................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.
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 thorough-
ly. Gently heat to boiling. Cool to room temperature. Add the  L-
cysteine·HCl·H20, 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 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.
Use:  For the cultivation and maintenance of Eubacterium lentum.