Citrobacter freundii|186116 |BNCC

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Citrobacter freundii

Subculture procedure	① Prepare 1-2 tablets mentioned above; ② After disinfecting the surface of the ampoule tube, open it in a 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 and transfer it into a freeze-drying tube. After fully dissolving the bacterial powder, transfer it into a Agar plate at a rate of 200 μ L per piece and apply evenly; ④ 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	1
morphology	The colony diameter is 1-2mm, circular, with irregular edges, opaque, yellow on the front, light color, raised in the middle, smooth surface, bright surface, and texture
Sharing mode	Public welfare sharing

Citrobacter freundii

Storage conditions: 2~8 ℃

No. 186116

Product format: freeze dried,200ul

Validity period: 6 years

Biosafety level: 1, handle in ultra-clear table or safety cabinet

Notes: 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.

Growth conditions:37 ℃, aerobic, nutritious agar/gravy medium (NA/NB). 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,pH7.0. 121 ℃, 15min.

Recovery steps:

(1)Prepare a flask of NB liquid media 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 18-24 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 fluid is turbid, NA plate colony is obvious
colony morphology: (above)	size: medium shape: round edge: neat edge transparency: translucent color: off-white uplift: uplift surface: smooth and bright texture: moist
conclusion:	good viability, and colony morphology is not abnormal, completely consistent with the above figure, qualified

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