Listeria monocytogenes|185986 |BNCC

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Listeria monocytogenes

Subculture procedure	① Prepare 1-2 of the above plates; ② Open the safety cabinet and follow the arrow indication on the cap of the penicillin bottle to open the strain and reconstitution solution; ③ Add 1mL of the complex solution to the Freeze dried pellet, cover with a rubber stopper, and shake to dissolve; ④ Take 200 μ L of bacterial suspension separately and inject it into two plates, apply evenly; ⑤ Place the petri dish under the above cultivation conditions for cultivation, and once the bacterial strain grows, it can be used.
Growth conditions	37 ℃; 18-24h; aerobic
Storage conditions	2-8 ℃
Safety level	2
morphology	The colony diameter is 1-2mm, circular, with neat edges, opaque, gray white on the front, raised in the middle, smooth on the surface, moist texture, and easy to pick up, G＋
Sharing mode	Public welfare sharing

Listeria monocytogenes

No.: 185986

Product format: freeze dried,200ul

Storage conditions: 2~8 ℃

Validity period: 6 years

Biosafety level: 2, handle in biosafety 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:37 ℃, aerobic, BHI: 4.0g of bovine brain extract powder, 4.0g of bovine heart extract powder, 5.0g of peptone, 16.0g of casin peptone, 5.0g of NaCl, 2.0g of glucose, 2.5g of disodium hydrogen phosphate, pH7.4±0.2. 121 ℃,15min.

Recovery steps:

(1)Prepare a flask of BHI liquid media or two BHI 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 BHI 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 24h BHI bacterial fluid is turbid, BHI plate colony is obvious
colony morphology:	size: smaller color: white shape: round edge: neat edge wet and dry: wet and smooth: smooth and bright transparency: opaque uplift: middle bulge
Conclusion:	good viability， and colony morphology is not abnormal, completely consistent with the above figure, qualified

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