Bacillus subtilis|186120 |BNCC

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Bacillus circulans

Subculture procedure	① Prepare 1-2 of the above plates; ② After disinfecting the surface of the ampoule tube, open it in a safety cabinet, burn the top with an alcohol lamp, quickly drip sterile water to break it, and then use tweezers to crush it; ③ Take 0.5mL of sterile water and transfer it into a freeze-drying tube. After fully dissolving the bacterial powder, transfer it into a Agar plate at a rate of 200 μ L per piece and apply evenly; ④ Place the petri dish under the above cultivation conditions for cultivation, and once the bacterial strain grows, it can be used.
Growth conditions	30 ℃; 18-24h; aerobic;
Storage conditions	2-8 ℃
Safety level	1
morphology	The colony diameter is 1-2mm, circular, with neat edges, opaque, gray white on the front, raised in the middle, smooth on the surface, moist texture, easy to pick up, G+(blue purple), Bacillus, purity: pure
Sharing mode	Public welfare sharing

Bacillus circulans

Storage conditions: 2~8 ℃

No. 186120

Product format: freeze dried,200ul

Validity period: 6 years

Biosafety level: 2, 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 plate 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℃， aerobic, nutrient agar medium. 18-24h, nutrient agar medium: beef extract 3.0g, peptone 10.0g, NaCl 5.0g, agar 20.0g (not included in liquid medium), distilled water 1.0L, pH 7.0. Sterilize at 121°C for 15min.

Recovery steps:

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

(2) Open the ampoule 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 results
viability	good viability，in 20h the liquid medium is turbid, and the plate bacteria layer is obvious; The bacterial liquid is marked on the plate, and the colony is typical
colony morphology: (above)	size: 1-2mm ; Shape: round ; Edge: irregular ; Transparency: opaque color: off-white ; Uplift: flat ; Surface: smooth and dark ; Texture: moist
Conclusion:	good viability, no abnormal colony morphology, qualified

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