Bacillus subtilis|192840 |BNCC

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Bacillus subtilis

Subculture procedure	① Prepare 1 test 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 shatter it; break into pieces with forceps; ③ Pipette 0.5 mL liquid medium into the freeze-dried vial, dissolve completely, then return to the liquid test tube and mix thoroughly; ④ Pipette 0.2mL of the bacterial suspension onto a Agar plate and spread evenly. 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	30°C; 18-24h; aerobic
Storage conditions	2-8°C
Safety level	1
morphology	Size: 1-2mm Shape: Circular Edge: Neat Transparency: Opaque Color: Pale yellow Elevation: Flat Surface: Smooth and shiny Texture: Moist and easily lifted
Sharing mode	Public welfare sharing

1.Description

1. Name:Bacillus subtilis

2. BNCC No.:192840

3. Biosafety level: 4

2.Storage conditions:

Storage of freezed dried ampoule and agar slant at 2°C to 8°C

3.Growth Conditions

1. Nutritious gravy agar: peptone 5.0g, beef extract 3.0g,NaCl 5.0g, agar 15.0g, distilled water 1.0L,pH7.0. [Note] Add 5mg MnSO4·H2O when culturing Bacillus，which is beneficial to spore production.

2. Atmosphere: aerobic

3. Temperature: 30 ℃,48h

4.Notes:

1.Normal culturing time, 24-48hours for bacterial, 72 hours for yeast, 5-7days for mould, 7-10days for fungal.

2.Agar slant shall be inoculated asap, and do not keep the storage for more than 3 months.

3.Please recover the strains in strict accordance with this instruction, otherwise the replacement of the strain are not be available in case of viability loss caused by different media or growth conditions.

4.Waste generated from the handling process should be discarded after high-pressure sterilization

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