Listeria monocytogenes|BNCC336877 |BNCC

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Listeria monocytogenes

Culture medium	Brain heart extract medium (BHIA/BHIB): brain extract 5.0g, heart extract 12.5g, peptone 5.0g, tryptone 10.0g, sodium chloride 5.0g, glucose 2.0g, disodium hydrogen phosphate 2.5g, agar 20.0g (not included in liquid medium),pH 7.4±0.2. Sterilization at 121 ℃ for 15min.
Subculture procedure	(1) Prepare a test tube containing 5~10mL of liquid medium and 2 plates; (2) Open it in the safety 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	37°C;18-24h; Aerobic
Growth characteristics	G Corynebacterium brevis, dynamic, catalase positive, β-hemolysis on the blood plate; negative nitrate reduction test, negative urease test, no fermentation of xylose and mannitol; positive methyl red reaction, positive Volt-Pu test, fermentation of glucose, maltose, heptaside, rhamnose; trisugar iron test produces acid, no hydrogen sulfide; facultative anaerobic, the most suitable pH7.4 ~ 7.6.
Storage conditions	2-8 ℃.
morphology	The colony diameter is 1-2mm, round, with neat edges, opaque, gray-white on the front and back, flat in shape, smooth in surface, bright in surface, moist in texture, easy to stir up, G + (blue-purple), bacilli, purity: pure
Separation substrate	Human
application	Research; Analytical testing. research, quality control. Quality control strain, "Quality Requirements for GB 4789.28 Medium and Reagents" standard strain.
Sharing mode	Public welfare sharing

Listeria monocytogenes

Storage conditions: 2~8 ℃

No.: 336877

Product format: freeze dried, 200ul

Validity period: Freeze-dried tube for 6 years

Biosafety level: 2 , handle in 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. 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 (without liquid medium), distilled water 1.0L,pH7.0. 121 ℃,15min.

Recovery steps:

(1)Prepare a flask of NB liquid media(fill 100ml triangulated bottles with 50ml medium) 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 20h NB bacterial liquid is turbid, obvious strain layer occurs on the plate; colony is typical on marked plate
colony morphology	size: general color: off-white shape: round edge: neat wet and dry: wet and smooth: smooth transparency: opaque uplift: uplift
conclusion	good viability, and colony morphology is not abnormal, completely consistent with the above figure, qualified