Entamoeba dispar Axenic Culture Medium

[所属分类:培养基配方] [发布时间:2020-7-7] [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

Entamoeba dispar Axenic
Culture Medium

培养基配方
Composition per liter:
YI-S solution.............................................................990.0mL
Gastric mucin stock suspension.............................................10.0mL
YI-S Solution:
Composition per liter:
YI broth..................................................................880.0mL
Bovine serum, heat inactivated............................................100.0mL
Vitamin mixture 18.........................................................20.0mL
Source:  Vitamin mixture 18 is available from Biofluids, Rockville, MD.
YI Broth:
Composition per liter:
YI base stock.............................................................780.0mL
10× Glucose buffer stock.................................................100.0mL
YI Base Stock:
Composition per 780.0mL:
Yeast extract...............................................................30.0g
L -Cysteine·HCl ............................................................1.0g
NaCl.........................................................................1.0g
Ascorbic acid ...............................................................0.2g
Ferric ammonium citrate...................................................228.0mg
10× Glucose Buffer Stock:
Composition per 100.0.0mL:
Glucose .....................................................................10.0g
K2HPO4 .......................................................................1.0g
KH2PO4 .......................................................................0.6g
Preparation of 10× Glucose Buffer Stock: Add components to
distilled/deionized water and bring volume to 100.0mL. Mix thorough-
ly. Filter sterilize.
Preparation of YI Base Stock: Add components to 600.0mL of
distilled/deionized water. Mix thoroughly. Bring volume to 780.0mL
with distilled/deionized water. Adjust pH to 6.8 with 1N NaOH. Dis-
tribute in 78.0mL aliquots to 100.0mL screw-capped bottles. Auto-
clave for 15 min at 15 psi pressure–121°C. Cool to room temperature.
Preparation of YI Broth: Aseptically add 10.0mL of 10× glucose
buffer stock to 78.0mL of cooled YI base stock. Adjust osmolarity with
NaCl to 380.0milliosmols/kg.
Preparation of YI-S Solution: Aseptically add 2.0mL of vitamin
mixture 18 and 10.0mL of heat-inactivated bovine serum to 88.0mL of
YI broth. Distribute in 12.0mL aliquots to 16 x 125mm screw-capped
test tubes. Store at 4°C in the dark with the caps screwed on tightly. Use
within 96 hr.
Gastric Mucin Stock Suspension:
Composition per 100.0mL:
Gastric mucin...............................................................1.0g
Preparation of Gastric Mucin Stock Suspension: Add gastric
mucin to distilled/deionized water and bring volume to 100.0mL. Mix
thoroughly. Autoclave for 15 min at 15 psi pressure–121°C.
Preparation of Medium:  Aseptically add 0.12mL of gastric mucin
stock suspension (thoroughly mix by swirling before use) to 12.0mL of
the complete YI-S solution in a 16 x 125mm screw-capped test tube.
Store at 4°C in the dark with the caps screwed on tightly. Use within 96
hrs.
Use: For the cultivation of Entamoeba dispar.