Bacillus subtilis|BNCC223758 |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
Growth characteristics	The round and shiny surface of the colony is smooth and the milky white edge is arched. The bacteria are arranged in a rod-like shape and move closely. Gram staining positive contact enzyme V-P determination hydrolyzed gelatin hydrolyzed starch-can grow in the range of 5 ℃-50 ℃. The salt tolerance range is wide, and it grows on beef paste medium containing 2-7% NaCl
Storage conditions	2-8 ℃
morphology	Size: 2-4mm Shape: Round Edge: False Filamentous Transparency: Opacity Color: off-white uplift: flat surface: rough gray texture: dry and easy to stir up
application	Special for livestock and poultry feed additives
Sharing mode	Public welfare sharing

Bacillus sublitis

BNCC No.：223758

Storage conditions: 2~8 ℃

Product format： freeze dried, 200ul

Validity period: 6 years

Biosafety level: 1, handle in ultra-clear table or safety cabinet

Receiving notice: if any abnormality is found on the day of receipt, please contact the customer service within 24 hours. If it is overdue, it is deemed that the bacteria are well. The freeze dried culture shall be used up once and shall not be retained. The bacterial viability will resume after 1-2 generation of recovery and can be used normally. Handling instructions of agar slant and suspension liquid is enclosed. Please operate in strict accordance with this instruction, otherwise the replacement of bacteria are not be available in case of aberrant growth and loss of viability.

Growth conditions:30°C, aerobic, nutrient agar medium, 18-24h. Nutritional agar medium: beef paste 3.0g, peptone 10.0g,NaCl 5.0g, agar 20.0g (not included in liquid medium), distilled water 1.0L,pH7.0. 121 ℃,15min.

Recovery steps:

(1)Prepare a test tube containing 5-10mL of liquid culture medium and 2 plates;

(2)Open it in the biosafety 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.

Recovery record: According to the recovery instructions, the results of the recovery are reported as follows:

item	test result
viability	good viability, in20h liquid medium become turbid, obvious strain layer occurs on the plate,colony is typical on marked plate.
colony morphology: (above)	size: 2-4mm shape: round edge: false filamentous transparency: opaque color: gray uplift: flat surface: rough gray texture: dry easy to pick up
conclusion:	good viability, no abnormal colony morphology, qualified