Bacillus subtilis subspecies|190344 |BNCC

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Bacillus subtilis subsp. subtilis

Subculture procedure	① Prepare one test tube containing 5-10 mL of liquid culture medium and two plates; ② Open the safety cabinet, burn the top with an alcohol lamp, quickly drip sterile water to break it, and then use tweezers to crush it; ③ Transfer 0.5mL of liquid culture medium into a freeze-drying tube, dissolve it thoroughly, and then transfer it back into the liquid test tube, mixing well; ④ Transfer 0.2mL of bacterial suspension into a Agar plate, apply evenly, and repeat twice to obtain two plates; ⑤ Place all liquid test tubes and plates under the above cultivation conditions, and the bacterial strains can be used once they grow.
Growth conditions	30 ℃; 18-24h; aerobic
Storage conditions	2-8 ℃
Safety level	1
morphology	Size: 4-6mm Shape: Circular Edge: Irregular Transparency: Opaque Color: Grayish-white Elevation: Flat Surface: Bright and rough Texture: Moist and viscous
Sharing mode	Public welfare sharing

Bacillus subtilis subsp. subtilis

BNCC No.:190344

Biosafety level: 4

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

Growth Conditions:

1.LB broth: yeast extract 5.0g, peptone 10.0g,NaCl 10.0g, distilled water 1.0L,pH 7.0. Sterilization at 121 ℃ for 15min.

2. Atmosphere: aerobic

3. Temperature: 30 ℃,48h

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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