BeNa Culture Collection
info@bncc.com
| Subculture procedure | ① Prepare 1 tube containing 5–10 mL liquid medium and 2 plates; ② Open in a safety cabinet, cauterize the top with an alcohol lamp, then quickly add sterile water to rupture it; break it into pieces with forceps; ③ Pipette 0.5mL liquid medium into the freeze-dried vial, dissolve thoroughly, then return to the liquid test tube and mix well; ④ Pipette 0.2mL bacterial suspensioninto each Agar plate, spread evenly, and repeat twice to obtain two plates; ⑤ Incubate all liquid tubes and plates under the specified conditions. The culture is ready for use once the bacterial strain grows. |
| Growth conditions | 37°C, 18-24h, aerobic |
| Storage conditions | 2-8°C |
| Safety level | 1 |
| morphology | Size: 2-4mm Shape: Irregular Edges: Pseudohyphal Transparency: Opaque Color: Grayish-white Elevation: Flat Surface: Rough and dull Texture: Dry and easily lifted |
| Sharing mode | Public welfare sharing |
Bacillus subtilis subsp.subtilis
No.: 231928
Storage conditions: 2~8 ℃
Product format: freeze dried,200ul
Validity period: 6 years
Biosafety level: 1, handle in ultra-clean table or safety cabinet
Receiving notice: if any abnormality is found on the day of receipt, please contact the customer service within 24 hours. If it is overdue, it is deemed that the bacteria are well. The freeze dried culture shall be used up once and shall not be retained. The bacterial viability will resume after 1-2 generation of recovery and can be used normally. Handling instructions of agar slant and suspension liquid is enclosed. Please operate in strict accordance with this instruction, otherwise the replacement of bacteria are not be available in case of aberrant growth and loss of viability.
Growth conditions:37 ℃, aerobic, LB agar medium, 18-24h. LB agar medium: yeast extract 5.0g, peptone 10.0g,NaCl 10.0g, agar 15.0g, distilled water 1.0 L,pH 7.0. 121 ℃,15min.
Recovery steps:
(1)Prepare a test tube containing 5-10mL of liquid culture medium and 2 plates;
(2) Open it in the biosafety cabinet, heat the tip of ampoule in a flame, quickly drop sterile water to creak it, then break it with forceps;
(3) Draw 0.5mL of liquid culture medium into a freeze dried ampoule, fully rehydrate and transfer the solution to the liquid test tube, mix evenly;
(4) Inoculate a plate with 0.2mL of suspension liquid, repeat the step to obtain two plates;
(5) Put all the liquid test tubes and plates under the above culture conditions for cultivation, and the strains can be used when they grow.
Recovery record: According to the recovery instructions, the results of the recovery are reported as follows:

| item | test result |
| viability: | good viability, in 20h liquid medium become turbid, obvious strain layer occurs on the plate,colony is typical on marked plate. |
|
colony morphology: (above) |
size: 2-4mm shape: irregular edge: false filamentous transparency: opaque color: gray uplift: flat surface: rough gray texture: dry easy to pick up |
| conclusion: | good viability, no abnormal colony morphology, qualified |