Citrobacter freundii|BNCC186116 |BNCC

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Citrobacter freundii

Culture medium	Nutritional gravy medium (NA/NB): 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. [Note] Adding 5 mg MnSO4 · H2O when culturing Bacillus. it is beneficial to produce spores.
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 ℃;,18-24h; Aerobic;
Storage conditions	2-8 ℃
morphology	The colony diameter is 1-2mm, round, with neat edges, opaque, yellow on the front, light color, convex in the middle, bright surface, smooth surface, moist texture, easy to pick up, G-(red), bacilli
application	API verification strain., "Quality Requirements for GB 4789.28 Medium and Reagents" Standard Strain
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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