ETSA Medium

[所属分类:培养基配方] [发布时间:2020-7-9] [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

ETSA Medium

培养基配方
Composition per 998.0mL:
Agar.......................................................................19.0g
Pancreatic digest of casein................................................15.0g
Papaic digest of soybean meal...............................................5.0g
NaCl........................................................................5.0g
Yeast extract...............................................................1.0g
KNO3 .......................................................................0.5g
Sodium formate .............................................................0.5g
Sodium succinate............................................................0.5g
Sheep blood, defibrinated.................................................30.0mL
L -Cysteine·HCl·H2O solution ...........................................10.0mL
Dithiothreitol solution...................................................10.0mL
Glucose solution .........................................................10.0mL
Na2CO3 solution...........................................................10.0mL
Menadione solution.........................................................2.0mL
Sodium fumarate solution...................................................2.0mL
Sodium lactate (60% syrup).................................................1.3mL
Hemin solution..............................................................1.0mL
pH 7.3 ± 0.2 at 25°C
Hemin Solution:
Composition per 200.0mL:
KOH.........................................................................1.12g
Hemin........................................................................0.2g
Ethanol (95% solution)....................................................100.0mL
Preparation of Hemin Solution:  Add KOH to distilled/deionized
water and bring volume to 100.0mL. Mix thoroughly. Add ethanol.
Mix thoroughly. Add hemin. Mix thoroughly.
Menadione Solution:
Composition per 100.0mL:
Menadione (vitamin K 3 )...................................................1.0g
Ethanol (95% solution)...................................................50.0mL
Preparation of Menadione Solution:  Add menadione to 50.0mL
of ethanol. Mix thoroughly. Bring volume to 100.0mL with distilled/
deionized water. Filter sterilize.
L -Cysteine·HCl·H2O Solution:
Composition per 10.0mL:
L- Cysteine·HCl·H2O.......................................................0.4g
Preparation of  L -Cysteine·HCl·H2O Solution: Add  L -cyste-
ine·HCl·H2O to distilled/deionized water and bring volume to 10.0mL.
Mix thoroughly. Filter sterilize.
Dithiothreitol Solution:
Composition per 10.0mL:
Dithiothreitol.............................................................0.05g
Preparation of Dithiothreitol Solution: Add dithiothreitol to
distilled/deionized water and bring volume to 10.0mL. Mix thorough-
ly. Filter sterilize.
Glucose Solution:
Composition per 10.0mL:
D- Glucose..................................................................1.0g
Preparation of Glucose Solution: Add glucose to distilled/deion-
ized water and bring volume to 10.0mL. Mix thoroughly. Filter steril-
ize.
Sodium Fumarate Solution:
Composition per 10.0mL:
Sodium fumarate...........................................................0.1g
Preparation of Sodium Fumarate Solution: Add sodium fu-
marate to distilled/deionized water and bring volume to 10.0mL. Mix
thoroughly. Filter sterilize.
Na2CO3 Solution:
Composition per 10.0mL:
Na2CO3 ...................................................................0.4g
Preparation of Na2CO3 Solution: Add Na2CO3 to distilled/deion-
ized water and bring volume to 10.0mL. Mix thoroughly. Filter steril-
ize.
Preparation of Medium: Add components—except menadione so-
lution,  L -cysteine·HCl·H2O solution, dithiothreitol solution, glucose
solution, sodium fumarate solution, Na2CO3 solution, and sheep
blood—to distilled/deionized water and bring volume to 926.0mL.
Mix thoroughly. Gently heat and bring to boiling. Autoclave for 15 min
at 15 psi pressure–121°C. Cool to 50°–60°C. Aseptically add in the fol-
lowing order: 2.0mL of sterile menadione solution, 10.0mL of sterile
L -cysteine·HCl·H2O solution, 10.0mL of sterile dithiothreitol solution,
10.0mL of sterile glucose solution, 2.0mL of sterile sodium fumarate
solution, 10.0mL of sterile Na2CO3 solution, and 30.0mL of sterile
sheep blood. Mix thoroughly. Aseptically and anaerobically distribute
into tubes under 80% N2 + 10% CO2 + 10% H2 . Cap tubes with butyl
rubber stoppers. 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 Bacteroides pneumosin-
tes, Falcivibrio grandis, Falcivibrio vaginalis, Mobiluncus curtisii,
Mobiluncus mulieris, Serpula hyodysenteriae, and Treponema species.