Thermophilic and lipophilic Bacillus subtilis|136223 |BNCC

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Geobacillus stearothermophilus

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	45 ℃, 18-24h, aerobic
Storage conditions	Freeze dried pellet
Safety level	1
morphology	Size: 1-2mm, shape: circular, edge: entire, transparency: opaque, color: grayish-white, elevation: flat, surface: glossy and smooth, texture: moist and easily picked up.
Sharing mode	Public welfare sharing

Geobacillus stearothermophilus

Storage conditions: 2~8 ℃

No. 136223

Product format: freeze dried,200ul

Validity period: 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. Handling instructions of agar slant and suspension liquid 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:45 ℃, aerobic, nutrient agar medium, 18-24h. 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,pH 7.0. Sterilization at 121 ℃ 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 24 hours,liquid medium become turbid, obvious strain layer occurs on the plate; colony is typical on marked plate
colony morphology: (above)	size: 1-2mm ; Shape: round ; Edge: neat ; Transparency: opaque color: off-white ; Protuberance: flat ; Surface: bright and smooth ; Texture: moist and easy to stir
Conclusion:	good viability, no abnormal colony morphology, qualified

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