Bacillus amyloliquefaciens|132867 |BNCC

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Bacillus amyloliquefaciens

Subculture procedure	① Prepare one test tube containing 5-10 mL of liquid culture medium and two 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 Transfer 0.5mL of liquid culture medium into a freeze-drying tube, dissolve it thoroughly, and then transfer it back into the liquid test tube, mixing well; ④ Transfer 0.2mL of bacterial suspension into a Agar plate, apply evenly, and repeat twice to obtain two plates; ⑤ Place all liquid test tubes and plates under the above cultivation conditions, and the bacterial strains can be used once they grow.
Growth conditions	30 ℃; 18-24h; aerobic
Storage conditions	2-8 ℃
Safety level	1
morphology	The colonies are 0.5-1 mm in diameter, irregular in shape with irregular margins, opaque, flat, and grayish-white on the surface. They have a rough, dry texture. The cells are Gram-positive (blue-violet) rods.
Sharing mode	Public welfare sharing

Bacillus subtilis subsp.

Storage conditions : 2~8 ℃

No.: 132867

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, nutrient 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 NA 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℃ (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 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 22h NB bacterial fluid become turbid, bacterial fluid marking MA plate marking is obvious
colony morphology	size: larger Color: yellowish Shape: round ; Edge: irregular edges Dry and wet: dry Smooth: rough opaque uplift: flat
conclusion	good viability, and colony morphology is not abnormal, completely consistent with the above figure, qualified

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