Bifidobacterium brevis|186529 |BNCC

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Bifidobacterium breve

Subculture procedure	① Prepare 1-2 fresh agar plates mentioned above (placed in an anaerobic environment for 24 hours of deoxygenation in advance); 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 (placed in an anaerobic environment for 24 hours of deoxygenation in advance) and pour it into a freeze-drying tube, dissolve it thoroughly, and mix well; ④ Transfer 0.2mL of bacterial suspension into a Agar plate, apply evenly, and repeat twice to obtain two plates; ⑤ Cultivate under the above conditions, and the strain can be used once it grows.
Growth conditions	37 ℃; 48-72h; anaerobic;
Storage conditions	2-8 ℃
Safety level	1
morphology	The colony diameter is 1-2mm, circular, with irregular edges, opaque, white on the front, raised in the middle, smooth on the surface, easy to pick up, G+(blue purple), Bacillus, purity: pure
Sharing mode	Public welfare sharing

Bifidobacterium breve

Storage conditions: 2~8 ℃

No. 186529

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 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: 37℃, anaerobic, BBL medium, 48-72h, BBL medium: peptone 15g, glucose 20g, yeast extract powder 2g, soluble starch 0.5g, sodium chloride 5g, L-cysteine 0.5g, tomato extract Powder 5g, liver extract powder 2g, Tween 80g, agar 20.0g (not included in liquid medium), distilled water 1.0L. pH 6.8±0.2. Sterilize at 121℃ for 15min.

Recovery steps:

① Prepare 1 test tube containing 5~10mL liquid medium and 2 plates (placed in an anaerobic environment to deoxygenate for 24h in advance)

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 72hours liquid medium become turbid,strain layer is obvious;colony is typical on the streaked plate
colony morphology: (above)	size: 0.5-1mm ; Shape: round ; Edge: irregular ; Transparency: opaque color: light yellow ; Protuberance: middle bulge ；Surface: rough ; Texture: dry is easy to provoke
Conclusion:	good viability, no abnormal colony morphology, qualified

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