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ONPG Broth

[所属分类:培养基配方] [发布时间:2021-12-7] [发布人:网站管理员2] [阅读次数:] [返回]

ONPG Broth

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

Composition per liter:

Peptone water.............................................. 750.0mL

ONPG solution............................................ 250.0mL

pH 7.2–7.4 at 25°C

ONPG Solution:

Composition per 250.0mL:

ONPG (o-Nitrophenyl-β-

D-galactopyranoside) ....................................1.5g

Sodium phosphate

buffer (0.01M, pH 7.5) ......................... 250.0mL

Preparation of ONPG Solution: Add ONPG to

250.0mL of sodium phosphate buffer. Mix thorough

ly. Filter sterilize.

Peptone Water:

Composition per 750.0mL:

Peptone ...............................................................7.5g

NaCl..................................................................3.75g

Preparation of Peptone Water: Add compo

nents to distilled/deionized water and bring volume

to 750.0mL. Mix thoroughly. Gently heat and bring

to boiling. Adjust pH to 8.0–8.4. Continue boiling for

10 min. Filter through Whatman #1 filter paper. Re

adjust pH of filtrate to 7.2–7.4. Autoclave for 20 min

at 10 psi pressure–115°C. Cool to 25°C.

Preparation of Medium: Aseptically combine

the sterile ONPG solution with the cooled, sterile

peptone water. Mix thoroughly. Aseptically distrib

ute into tubes in 2.5–3.0.0mL volumes. Store at 4°C

for up to 1 month.

Use: For the differentiation of a variety of Gram

negative bacteria based on production of β-galactosi

dase. For the differentiation of lactose-delayed bacte

ria from lactose-negative bacteria. For the differenti

ation of Pseudomonas cepacia (positive) and

Pseudomonas maltophila (positive) from other

Pseudomonas species (negative). Bacteria that pro

duce β-galactosidase turn the medium yellow.