Kingella kingae|BNCC357570 |BNCC

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Kingella kingae

Culture medium	Columbia blood plate (CA-B): casein pancreatin digest 10.0g, cardiopancreatin digest 3.0g, corn starch 1.0g, meat gastric enzyme digest 5.0g, yeast extract 5.0g, sodium chloride 5.0, agar 20.0g, distilled water 1.0L,pH 7.3±0.2. After sterilization at 121 ℃ for 15min, cool to 55 ℃, add 5% sterile defibrated sheep blood, shake well, pour the plate and cool it for later use.
Subculture procedure	(1) Prepare a test tube containing 5~10mL of liquid medium and 2 plates; (2) open it in the biosafety cabinet, heat the tip of ampoule in the flame, quickly drop sterile water to creak it, and 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	Size: 1-2mm Shape: Round Edge: Neat Transparency: Opaque Color: off-white uplift: flat surface: bright and smooth texture: moist and easy to stir up
Separation substrate	The intestines of pigs
Sharing mode	Public welfare sharing

Kingella kingae

Storage conditions : 2~8 ℃

No. : 357570

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 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, 18-24h. Columbia blood plate (ready-to-use) Brain Heart Infusion Medium (BHI): Brain Infusion Powder 5.0g, Heart Infusion Powder 12.5g, Peptone 5.0g, Tryptone 10.0g, Sodium Chloride 5.0g, Glucose 2.0g, Hydrogen Phosphate Disodium 2.5g, agar 20.0g (not included in liquid medium), pH 7.4±0.2. Sterilize at 121°C for 15min.

Recovery steps:

(1) Prepare a test tube containing 5-10mL of liquid culture medium and 2 plates;

(2) Open it 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 18-24h liquid medium become turbid, obvious strain layer occurs on the plate; colony is typical on marked plate
colony morphology (above)	size: 1-2mm shape: round edge: neat transparency: opaque color: off-white uplift: flat surface: bright and smooth texture: moist and easy to stir
conclusion	good viability, no abnormal colony morphology, qualified

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