H Broth (Hominis Broth)

[所属分类:培养基配方] [发布时间:2021-3-23] [发布人: ] [阅读次数:] [返回]

H Broth

(Hominis Broth)

培养基配方

Composition per 99.0mL:
Base broth................................................................................65.0mL
Horse serum.............................................................................20.0mL
Yeast dialysate..........................................................................10.0mL
Penicillin solution......................................................................2.0mL
Glucose solution ........................................................................1.0mL
Thallium acetate solution...........................................................1.0mL
pH 7.3 ± 0.2 at 25°C
Base Broth:
Composition per liter:
Papaic digest of soybean meal....................................................20.0g
NaCl..............................................................................................5.0g
Phenol Red (2% solution)..........................................................1.0mL
Preparation of Base Broth: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently heat and bring to boiling. Adjust pH to 7.3. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 25°C.
Yeast Dialysate:
Composition per 10.0mL:
Dried yeast, active.....................................................................450.0g
Preparation of Yeast Dialysate: Add active, dried yeast to distilled/deionized water and bring volume to 1250.0mL. Gently heat and bring to 40°C. Autoclave for 15 min at 15 psi pressure–121°C. Put into dialysis tubing. Dialyze against 1.0L of distilled/deionized water for 2 days at 4°C. Discard tubing and its contents. Autoclave dialysate for 15 min at 15 psi pressure–121°C. Store at −20°C.
Penicillin Solution:
Composition per 10.0mL:
Penicillin..............................................................................100,000U
Preparation of Penicillin Solution: Add penicillin to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize.
Glucose Solution:
Composition per 10.0mL:

D- Glucose......................................................................................1.8g

Preparation of Glucose Solution: Add  D- glucose to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize.
Thallium Acetate Solution:
Composition per 10.0mL:
Thallium acetate..........................................................................0.33g
Preparation of Thallium Acetate Solution: Add thallium acetate to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize.
Caution: Thallium salts are toxic.
Preparation of Medium: To 65.0mL of cooled, sterile base broth, aseptically add 10.0mL of sterile yeast dialysate, 20.0mL of horse serum, 2.0mL of sterile penicillin solution, 1.0mL of sterile glucose solution, and 1.0mL of sterile thallium acetate solution. Mix thoroughly. Aseptically distribute into sterile screw-capped tubes in 5.0mL volumes. Screw caps down tightly.
Use: For the isolation and cultivation of Mycoplasma pneumoniae.