Bacillus subtilis|BNCC188159 |BNCC

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Bacillus subtilis

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 when culturing Bacillus. it 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: 1-2mm Shape: Round Edge: Irregular Transparency: Opaque Color: off-white bulge: middle bulge surface: rough fold texture: wet and easy to stir up
application	Lipase production.
Sharing mode	Public welfare sharing

Bacillus subtilis

BNCC No.:188159

Biosafety level: 4

Storage conditions:

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

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: 28-30 ℃

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.