Bacillus sphaericus|BNCC193424 |BNCC

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Bacillus sphaericus

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	37 ℃;18-24h; Aerobic
Storage conditions	2-8 ℃
morphology	Size: 4-6mm Shape: Round Edge: Neat Transparency: Opaque Color: off-white uplift: flat surface: bright and smooth texture: moist and easy to stir up
Sharing mode	Public welfare sharing

Bacillus sphaericus

No.: 193424

Storage conditions: 2~8 ℃

Product format： freeze dried, 200ul

Validity period: Freeze-dried tube for 24 months

Biosafety level: 1, handle in ultra-clean table or biosafety 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:37 ℃, aerobic, nutritious agar/gravy medium (NA/NB). 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 flask of NB liquid media or two NB agar plates.

(2)Open it in 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 medium into the freeze dried ampoule,make the strain pellet fully dissolved. transfer the liquid suspension to the liquid media, and culture in plate shaker at 37℃ for 18-24 hours (140r/min); or directly dispense 200ul of the liquid suspension into a NA agar plate evenly, then put the plates in incubator at 37℃ for 24-48 hours.

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

item	test results
Viability:	good viability,in 20h NB bacterial fluid is turbid, NA plate colony is obvious
colony morphology:	size: larger color: orange-white shape: round edge: neat edge wet and dry: slightly dry and smooth: dark transparency: opaque uplift: flat, low convex
conclusion:	good viability, and colony morphology is not abnormal, completely consistent with the above figure, qualified