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Entamoeba dispar Axenic Culture Medium

[所属分类:培养基配方] [发布时间:2021-11-17] [发布人:网站管理员2] [阅读次数:] [返回]

Entamoeba dispar Axenic

Culture Medium

山东拓普生物工程有限公司培养基配方 http://www.topbiol.com

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 Bio

fluids, Inc., 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 thoroughly. Filter

sterilize.

Preparation of YI Base Stock: Add components

to 600.0mL of distilled/deionized water. Mix thor

oughly. Bring volume to 780.0mL with distilled/

deionized water. Adjust pH to 6.8 with 1N NaOH.

Distribute in 78.0mL aliquots to 100.0mL screw

capped bottles. Autoclave for 15 min at 15 psi pres

sure–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-in

activated bovine serum to 88.0mL of YI broth. Dis

tribute 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 Suspen

sion: Add gastric mucin to distilled/deionized water

and bring volume to 100.0mL. Mix thoroughly. Au

toclave for 15 min at 15 psi pressure–121°C.

Preparation of Medium: Aseptically add

0.12mL of gastric mucin stock suspension (thor

oughly 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 hr.

Use: For the cultivation of Entamoeba dispar.

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Entamoeba dispar Axenic Culture Medium