Leptospira Protein-Free Medium (Leptospira PF Medium)

[所属分类:培养基配方] [发布时间:2021-4-30] [发布人:李佳颐] [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
MgCl 2 -CaCl 2 solution................................................................1.0mL
Cyanocobalamin (0.02% solution) ............................................1.0mL
Glycerol (10% solution) ............................................................1.0mL
KH 2 PO 4 (1% solution)...............................................................1.0mL
MnSO 4 ·H 2 O (0.1% solution).....................................................1.0mL
ZnSO 4 (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 thoroughly. While stirring, add charcoal.
Stir mixture for 18 hr at 25°C. Allow charcoal to settle out of suspen-
sion 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. De-
cant supernatant solution. Pass Tween™ solution through a thin-chan-
nel 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 thoroughly. While stirring, add charcoal.
Stir mixture for 18 hr at 25°C. Allow charcoal to settle out of suspen-
sion 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. De-
cant supernatant solution. Pass Tween™ solution through a thin-chan-
nel ultrafiltration XM 100 membrane. Store stock solution at −20°C.
MgCl 2 -CaCl 2 Solution:
Composition per 100.0mL:
CaCl 2 ·2H 2 O ..................................................................................1.5g
MgCl 2 ·6H 2 O .................................................................................1.5g
Preparation of MgCl 2 -CaCl 2 Solution: Add components to dis-
tilled/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. Asep-
tically distribute into sterile tubes or flasks.
Use: For the cultivation of Leptospira species.