BeNa Culture Collection
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| Culture medium | BNCC352241 |
| Description | Columbia blood agar plate |
| Composition | Special Peptone:23.0g, Soluble Starch:1.0g, NaCl:5.0g, Agar:10.0g, Defibrinated Sheep Blood:5%(added at 50℃), pH:7.3±0.2(25℃) |
| Growth conditions | 37°C; 4-5 days; microaerophilic (3%-5% O₂, 10% CO₂); |
| Subculture procedure | ① Thaw Frozen vials in a 37°C water bath with vigorous shaking; ② Disinfect outer surface of vial with alcohol swab, then transfer to safety cabinet for handling; ③ Unscrew cap and aseptically transfer entire thawed culture to 5-6 mL brain heart infusate broth tube. ④ Inoculate 0.5 mL broth culture onto Agar slant; pour remaining culture onto Columbia blood agar plates and spread evenly. To achieve biphasic growth, loosely cap the slants and incubate at an angle. Gas supply is provided by a 2.5 L microaerophilic gas generator. Precautions: a. Use fresh, moist Columbia blood agar slants and plates; b. Biphasic growth on slants and plates should appear within 4-5 days, though turbidity is minimal. Growth can be observed via Gram staining of the broth; c. Upon obtaining robust growth, add equal volumes of 20% sterile glycerol to pooled broth from several biphasic slants, then freeze in Liquid nitrogen or at -80°C. |
| Storage conditions | -80°C |
| Safety level | 1 |
| morphology | G− (red), spirochetes or cocci, Purity: Pure |
| Sharing mode | Public welfare sharing |
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