Bacillus subtilis|194629 |BNCC

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Bacillus subtilis subsp. subtilis

Subculture procedure	① Prepare two plates as above; ② Disinfect ampoule surface, open in safety cabinet, cauterize top with alcohol lamp, then promptly add sterile water to rupture. Crush ampoule with forceps; ③ Pipette 0.5 mL sterile water into lyophilized tube. Thoroughly dissolve bacterial powder, then inoculate two plates (200 μL/Agar plate) with uniform spread; ④ Incubate plates under specified conditions until colonies emerge.
Growth conditions	30°C; 18-24h; aerobic
Storage conditions	2-8°C
Safety level	1
morphology	Colony diameter: 1-2 mm; irregular shape with jagged edges; opaque, pale yellow surface; flat morphology; rough texture; dry appearance; Gram-positive (blue-violet staining); bacilli
Sharing mode	Public welfare sharing

1.Description

1. Name：Bacillus subtilis subsp. subtilis

2. BNCC No.：194629

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: 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：37 ℃

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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