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培养基配方

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TYI-S-33 Medium

[所属分类:培养基配方] [发布时间:2021-9-29] [发布人:网站管理员2] [阅读次数:] [返回]

TYI-S-33 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:

Pancreatic digest of casein......................................................... 20.0g

Yeast extract................................................................................ 10.0g

NaCl .............................................................................................. 2.0g

L-Cysteine·HCl.............................................................................. 1.0g

Ascorbic acid ............................................................................... 0.2g

Ferric ammonium citrate..........................................................22.8mg

Bovine serum, heat inactivated..............................................100.0mL

Special 107 vitamin mix ........................................................100.0mL

Buffer solution .........................................................................50.0mL

Glucose solution ......................................................................50.0mL

pH 6.8 ± 0.2 at 25°C

Glucose Solution:

Composition per 50.0mL:

Glucose ....................................................................................... 10.0g

Preparation of Glucose Solution: Add glucose to distilled/deion

ized water and bring volume to 50.0mL. Mix thoroughly. Filter steril

ize.

Buffer Solution:

Composition per 50.0mL:

K2HPO4......................................................................................... 1.0g

KH2PO4........................................................................................ 0.6g

Preparation of Buffer Solution: Add components to distilled/de

ionized water and bring volume to 50.0mL. Mix thoroughly. Adjust pH

to 6.8. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C.

Special 107 Vitamin Mix:

Composition per 120.0.0mL:

Solution 1...................................................................................1.2mL

Solution 2...................................................................................0.4mL

Solution 3...................................................................................0.4mL

Solution 4 vitamins................................................................100.0mL

Preparation of Special 107 Vitamin Mix: Aseptically combine

1.2mL of sterile solution 1, 0.4mL of sterile solution 2, 0.4mL of sterile

solution 3, and 100.0mL of sterile solution 4 vitamins. Bring volume

to 120.0mL with sterile distilled/deionized water.

Solution 1:

Composition per 100.0mL:

Vitamin B12 .............................................................................. 40.0mg

Preparation of Solution 1: Add vitamin B12 to distilled/deionized

water and bring volume to 100.0mL. Mix thoroughly. Filter sterilize.

Solution 2:

Composition per 100.0.0mL:

DL-6,8-Thioctic acid, oxidized............................................... 100.0mg

Absolute ethanol ....................................................................100.0mL

Preparation of Solution 2: Add DL-6,8-thioctic acid to absolute

ethanol and bring volume to 100.0mL. Mix thoroughly. Filter sterilize.

Solution 3:

Composition per 100.0mL:

Tween™ 80................................................................................. 50.0g

Absolute ethanol ....................................................................100.0mL

Preparation of Solution 3: Add Tween™ 80 to absolute ethanol

and bring volume to 100.0mL. Mix thoroughly. Filter sterilize.

Solution 4 Vitamins:

Composition per liter:

Vitamin B12 .............................................................................. 10.0mg

Calcium D-(+)-pantothenate ...................................................... 2.3mg

Choline chloride....................................................................... 1.25mg

Riboflavin .................................................................................. 0.7mg

Calciferol (vitamin D2) ............................................................ 0.25mg

Vitamin A, crystallized alcohol ............................................... 0.25mg

p-Aminobenzoic acid............................................................. 0.125mg

i-Inositol................................................................................. 0.125mg

Biotin ..................................................................................... 0.025mg

α-Tocopherol phosphate, disodium salt................................. 0.025mg

Folic acid ............................................................................... 0.025mg

Menadione (vitamin K3) ........................................................ 0.025mg

Thiamine·HCl ........................................................................ 0.025mg

Niacin................................................................................... 0.0625mg

Niacinamide......................................................................... 0.0625mg

Pyridoxal·HCl ...................................................................... 0.0625mg

Pyridoxine·HCl .................................................................... 0.0625mg

Preparation of Solution 4 Vitamins: Add components to dis

tilled/deionized water and bring volume to 1.0L. Mix thoroughly. Filter

sterilize.

Preparation of Medium: Add components, except heat-inactivated

bovine serum, special 107 vitamin mix, glucose solution, and buffer

solution, to distilled/deionized water and bring volume to 700.0mL.

Mix thoroughly. Autoclave for 15 min at 15 psi pressure–121°C. Asep

tically add 100.0mL of sterile, heat-inactivated bovine serum, 50.0mL

of sterile glucose solution, 100.0mL of sterile special 107 vitamin mix,

and 50.0mL of sterile buffer solution. Mix thoroughly. Aseptically dis

tribute into sterile tubes or flasks.

Use: For the cultivation of Entamoeba barreti, Entamoeba histolytica,

Entamoeba insolita, Entamoeba invadens, Entamoeba moshkovskii,

Entamoeba ranarum, Entamoeba terrapinae, Monocercomonas spe

cies, Phreatamoeba balamuthi, Spironucleus vortens, and Trichomo

nas tenax.

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TYI-S-33 Medium