Bacillus subtilis|133158 |BNCC

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Bacillus thuringiensis

Subculture procedure	① Prepare 1 test tube containing 5–10 mL liquid medium and 2 plates; ② Open in a safety cabinet, cauterize the top with an alcohol lamp, then quickly add sterile water to break it; subsequently crush it with forceps; ③ Pipette 0.5 mL liquid medium into the freeze-dried vial, dissolve thoroughly, then return to the liquid test tube and mix well; ④ Pipette 0.2 mL of the bacterial suspension onto a Agar plate and spread evenly. Repeat twice to obtain two plates; ⑤ Incubate all liquid tubes and plates under the specified conditions. The culture is ready for use once bacterial growth appears.
Growth conditions	37°C, 24-48h, aerobic
Storage conditions	2-8°C
Safety level	1
morphology	Size: Large Color: Creamy white Shape: Circular Edge: Irregular Moisture: Dry Smoothness: Rough Transparency: Opaque Elevation: Flat
Sharing mode	Public welfare sharing

Bacillus thuringiensis

Storage conditions: 2~8 ℃

No. : 133158

Product format : freeze dried,200ul

Validity period: freezed-dried tube for 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. 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 :37°C, aerobic, nutrient gravy medium. 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. Activation steps:

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℃ 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 result
viability	good viability, in 40h NB bacterial fluid become turbid, bacterial fluid scribing plate colonies are obviously typical
colony morphology:	size: large color: milky white shape: round edge: irregular wet and dry: dry and smooth: rough transparency: opaque uplift: flat
Conclusion:	good viability, and colony morphology is not abnormal, completely consistent with the above figure, qualified

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