Bacillus subtilis(Ehrenberg)Cohn|BNCC336704 |BNCC

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Bacillus subtilis(Ehrenberg)Cohn

Culture medium	Nutritional gravy medium (NA/NB): beef paste 3.0g, peptone 10.0g,NaCl 5.0g, agar 20.0g (not included in liquid medium), distilled water 1.0L,pH 7.0. Sterilization at 121 ℃ for 15min. [Note] Adding 5 mg MnSO4 · H2O to culture Bacillus is beneficial to produce spores.
Subculture procedure	(1) Prepare a test tube containing 5~10mL of liquid medium and 2 plates; (2) Open it in the safety 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.
Growth conditions	30 ℃;18-24h; Aerobic
Storage conditions	2-8 ℃
morphology	Size: 2-4mm shape: round edge: neat transparency: opaque Color: light yellow uplift: flat surface: bright and smooth texture: moist and easy to stir up
Sharing mode	Public welfare sharing

1.Description

1. Name: Bacillussubtilis

2. BNCC No.:336704

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: beef paste 3.0g, peptone 10.0g,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. pH7.0. 121 ℃,15min.

2. Atmosphere: aerobic

3. Temperature: 30 ℃

4.Notes:

1.Normal culturing time, 1-2days for bacterial, 3 days for yeast, 5-7days for mould, 7-10days for fungal.

2.Agar slant shall be inoculated asap, and long-term storage is not recommended.

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.Users should ensure the safe storage and operation of the strain,waste generated from the handling process should be discarded after high-pressure sterilization.

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