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Leptospira Protein-Free Medium (Leptospira PF Medium)

[所属分类:培养基配方] [发布时间:2021-11-26] [发布人:网站管理员2] [阅读次数:] [返回]

Leptospira Protein-Free Medium

(Leptospira PF 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:

TES (N-Tris[hydroxymethyl]methyl-2-aminoethane

sulfonic acid) buffer .....................................1.2g

NaCl....................................................................0.9g

Sodium pyruvate.................................................0.2g

CT-Tween 60................................................. 12.0mL

CT-Tween 40................................................... 3.0mL

MgCl2-CaCl2 solution..................................... 1.0mL

Cyanocobalamin (0.02% solution) ................. 1.0mL

Glycerol (10% solution).................................. 1.0mL

KH2PO4 (1% solution).................................... 1.0mL

MnSO4·H2O (0.1% solution) .......................... 1.0mL

ZnSO4 (0.4% solution).................................... 0.1mL

pH 7.6 ± 0.2 at 25°C

CT-Tween 60:

Composition per 200.0mL:

Charcoal, Norit A..............................................40.0g

Tween 60...........................................................20.0g

Preparation of CT-Tween 60: Add Tween 60 to

200.0mL of distilled/deionized water. Mix thorough

ly. While stirring, add charcoal. Stir mixture for 18 hr

at 25°C. Allow charcoal to settle out of suspension

for 18 hr at 4°C. Carefully decant the Tween solution

off the sediment. Centrifuge the Tween solution at

10,000 × g for 1 hr. Decant supernatant solution. Pass

Tween solution through a thin-channel ultrafiltration

XM 100 membrane. Store stock solution at −20°C.

CT-Tween 40:

Composition per 200.0mL:

Charcoal, Norit A..............................................40.0g

Tween 40...........................................................20.0g

Preparation of CT-Tween 40: Add Tween 40 to

200.0mL of distilled/deionized water. Mix thorough

ly. While stirring, add charcoal. Stir mixture for 18 hr

at 25°C. Allow charcoal to settle out of suspension

for 18 hr at 4°C. Carefully decant the Tween solution

off the sediment. Centrifuge the Tween solution at

10,000 × g for 1 hr. Decant supernatant solution. Pass

Tween solution through a thin-channel ultrafiltration

XM 100 membrane. Store stock solution at −20°C.

MgCl2–CaCl2 Solution:

Composition per 100.0mL:

CaCl2·2H2O ........................................................1.5g

MgCl2·6H2O .......................................................1.5g

Preparation of MgCl2-CaCl2 Solution: Add

components to distilled/deionized water and bring

volume to 100.0mL. Mix thoroughly.

Preparation of Medium: Add components to

distilled/deionized water and bring volume to 1.0L.

Mix thoroughly. Filter sterilize. Aseptically distrib

ute into sterile tubes or flasks.

Use: For the cultivation of Leptospira species.

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Leptospira Protein-Free Medium (Leptospira PF Medium)