Salt denitrifying Bacillus subtilis|132612 |BNCC

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Virgibacillus halodenitrificans

Subculture procedure	① Prepare 1-2 of the above plates; ② Open the 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 two plates, each Agar plate containing about 200 μ L, 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 0.5-1mm, circular, with neat edges, opaque, yellow on the front, raised in the middle, smooth and bright on the surface, moist texture, easy to pick up, G+(blue purple), Bacillus, purity: pure
Sharing mode	Public welfare sharing

Virgibacillus halodenitrificans

Storage conditions : 2~8 ℃

No. 132612

Product format: freeze dried,200ul

Validity period: Freeze-dried tube for 24 months

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. 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.

Recovery/subculture :30 ℃, aerobic, 5% sodium chloride agar medium. 5% sodium chloride agar medium: beef paste 3.0g, peptone 10.0g,NaCl 50.0g, agar 20.0g (not included in liquid medium), distilled water 1.0L,pH7.0. 121 ℃,15min.

Recovery steps :

(1)Prepare 1 bottle of 5% sodium chloride liquid culture medium (100ml triangular bottle is divided into 50ml culture medium);

(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℃ (140r/min) for 18-24 hours.

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

item	test result
viability	good viability, in 20h,5% sodium chloride liquid medium become turbid
colony morphology:	size: medium color: light yellow shape: round edge: irregular edge wet and dry: dry and smooth: rough, dim transparency: opaque uplift: flat
Conclusion:	good viability, and colony morphology is not abnormal, completely consistent with the above figure, qualified

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