NOS Spirochete Medium

[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 Spirochete Medium

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

Composition per 1045.0mL:

Basal medium ...............................................................................1.0L
NaHCO 3 (10% solution)..........................................................20.0mL
Rabbit serum, heat inactivated.................................................20.0mL
Thiamine pyrophosphate (0.2% solution)..................................3.0mL
VFA solution..............................................................................2.0mL
pH 7.4 ± 0.2 at 25°C
Basal Medium:
Composition per liter:
Pancreatic digest of casein..........................................................10.0g
Pancreatic digest of gelatin.........................................................4.85g
Noble agar.....................................................................................3.0g
Yeast extract..................................................................................2.5g
Brain heart, solids from infusion..................................................2.0g
Peptic digest of animal tissue .......................................................2.0g
Glucose.........................................................................................2.0g
NaCl............................................................................................1.65g
Glucose.........................................................................................1.0g
L- Cysteine·HCl·H 2 O......................................................................1.0g
Na 2 HPO 4 .....................................................................................0.85g
Sodium thioglycolate....................................................................0.5g
L- Asparagine...............................................................................0.25g
Preparation of Basal Medium: Add components to distilled/de-
ionized water and bring volume to 1.0L. Mix thoroughly. Gently heat
and bring to boiling. Gas under O 2 -free 80% N 2  + 10% CO 2 + 10% H 2 .
Stopper and wire flask closed. Autoclave for 20 min at 15 psi pressure–
121°C. Cool to 45°–50°C.
VFA Solution:
Composition per 100.0mL:
KOH (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 KOH solution. Mix thoroughly. Filter sterilize. Store at 4°C.
Preparation of Medium: Combine 20.0mL of NaHCO 3 solution,
20.0mL of rabbit serum, 3.0mL of thiamine pyrophosphate solution,
and 2.0mL of VFA solution. Mix thoroughly. Filter sterilize. Open the
flask containing 1.0L of cooled, sterile basal medium while flushing
with O 2 -free 85% N 2  + 10% CO 2 + 5% H 2 . Aseptically add the filter-
sterilized mixture. Mix thoroughly. Aseptically and anaerobically dis-
tribute into sterile tubes or flasks.
Use:  For the cultivation and maintenance of Treponema denticola and
Treponema socranskii.