Bacillus sp|BNCC357972 |BNCC

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Bacillus sp

Culture medium	Nutritional gravy medium (NA/NB): beef paste 3.0g, peptone 10.0g,NaCl 5.0g, agar 20.0g (without liquid medium), distilled water 1.0L,pH 7.0. Sterilization at 121 ℃ for 15min. [Note] Adding 5 mg MnSO4 · H2O to culture Bacillus 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 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.
Growth conditions	30°C;18-24; Aerobic;
Storage conditions	2-8 ℃
morphology	Size: 1-2mm Shape: Round Edge: Irregular Transparency: Opaque Color: white uplift: flat surface: rough texture: dry
Sharing mode	Public welfare sharing

Bacillus sp

Storage conditions : 2~8 ℃

No. : 357972

Product format :freeze dried, 200ul

Validity : 6 years

Biosafety level : 1, handle in ultra-clean 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°C, aerobic, nutrient agar medium, 18-24h.

Nutrient agar medium: beef extract 3.0g, peptone 10.0g, NaCl 5.0g, agar 20.0g (liquid medium not included), distilled water 1.0L, pH 7.0. Sterilize at 121°C for 15min.

Recovery steps:

① Prepare 1 test tube containing 5~10mL liquid medium and 2 plates;

② Open it in the safety cabinet, burn the top with an alcohol lamp, then quickly drip sterile water to break it, and then smash it with tweezers;

③ Pipette 0.5mL of liquid medium into the freeze-drying tube, dissolve it fully and pour it back into the liquid test tube, and mix well;

④ Pipette 0.2mL of bacterial suspension into the plate, spread evenly, repeat twice to obtain two plates;

⑤ Put all the liquid test tubes and plates under the above-mentioned culture conditions, and the bacteria 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 24 hours liquid medium become turbid,strain layer is obvious;colony is typical on the streaked plate
colony morphology (above)	size: 1-2mm shape: round edge: irregular transparency: opaque color: white uplift: flat surface: rough texture: dry
Conclusion	good resurrection, no abnormal colony morphology, qualified

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