NOS Medium, Modified

[所属分类:培养基配方] [发布时间:2021-6-21] [发布人:网站管理员2] [阅读次数:] [返回]

NOS Medium, Modified

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

Composition per 100.67mL:

Basal medium..........................................................................94.0mL
NaHCO 3 solution.....................................................................2.67mL
TPP/VFA mixture......................................................................2.0mL
Rabbit serum, heat inactivated...................................................2.0mL
pH 7.4 ± 0.2 at 25°C
Basal Medium:
Composition per 94.0mL:
Pancreatic digest of casein............................................................1.0g
Pancreatic digest of gelatin.........................................................0.48g
Yeast extract................................................................................0.25g
Brain heart, solids from infusion..................................................0.2g
Peptic digest of animal tissue .......................................................0.2g
D- Glucose......................................................................................0.2g
NaCl............................................................................................0.17g
Glucose .........................................................................................0.1g
L- Cysteine·HCl·H 2 O......................................................................0.1g
Na 2 HPO 4 ...................................................................................0.085g
Sodium thioglycolate..................................................................0.05g
L- Asparagine .............................................................................0.025g
Resazurin (0.1% w/v solution)...................................................0.1mL
Preparation of Basal Medium: Add components to distilled/de-
ionized water and bring volume to 94.0mL. Mix thoroughly. Gently
heat and bring to boiling. Gas under O 2 -free 85% N 2  + 10% CO 2 + 5%
H 2 . Stopper and wire flask closed. Autoclave for 20 min at 15 psi pres-
sure–121°C. Cool to 45°–50°C.
NaHCO 3 Solution:
Composition per 10.0mL:
NaHCO 3 ......................................................................................0.75g
Preparation of NaHCO 3 Solution: Add the NaHCO 3 to distilled/
deionized water and bring volume to 10.0mL. Mix thoroughly. Filter
sterilize.
TPP/VFA Mixture:
Composition per 10.9mL:
Thiamine pyrophosphate (0.2% solution)..................................1.5mL
VFA solution..............................................................................1.0mL
Preparation of TPP/VFA Mixture: Add components to distilled/
deionized water and bring volume to 10.9mL. Mix thoroughly. Filter
sterilize. Store at −20°C.
VFA Solution:
Composition per 100.0mL:
NaOH (0.1N solution)..............................................................98.0mL
Isobutyric acid............................................................................0.5mL
2-Methylbutyric acid..................................................................0.5mL
Isovaleric acid............................................................................0.5mL
Valeric acid ................................................................................0.5mL
Preparation of VFA Solution: Add volatile fatty acids to 98.0mL
of NaOH solution. Mix thoroughly. Filter sterilize. Store at 4°C.
Preparation of Medium: Open the flask containing 94.0mL of
cooled sterile basal medium while flushing with O 2 -free 85% N 2  + 10%
CO 2 + 5% H 2 . Aseptically add sterile NaHCO 3 solution, sterile TPP/
VFA mixture, and filter-sterilized rabbit serum. Mix thoroughly.
Use:  For the cultivation and maintenance of Treponema vincentii and
other Treponema species.