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Leptospira Protein-Free Medium (Leptospira PF Medium)

[所屬分類:培養基配方] [發布時間:2021-11-26] [發布人:網站管理員2] [閱讀次數:] [返回]
Leptospira Protein-Free Medium
(Leptospira PF Medium)

山東拓普生物工程有限公司 培養基配方 //ljxinglong.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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