Klebsiella pneumoniae subsp.|BNCC358232 |BNCC

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Klebsiella pneumoniae subsp.

Culture medium	Tryptone soybean blood plate (TSA-B): tryptone 15.0g, soybean peptone 5.0g, sodium chloride 5.0g, 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 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	30°C;18-24h; Aerobic;
Storage conditions	2-8 ℃
morphology	Size: 2-4mm Shape: Round Edge: Irregular Transparency: Opaque Color: white bulge: middle bulge surface: bright and smooth texture: moist and easy to stir up
Separation substrate	Human urine, 59-yr-old male(hospital care in India)
application	Carbapenem resistant, NDM-1, MLST type ST14 (depositor)
Sharing mode	Public welfare sharing

Klebsiella pneumoniae subsp. pneumoniae

Storage conditions : 2~8 ℃

No. : 358232

Product format :freeze dried, 200ul

Validity : 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 and can be used normally. Handling instructions of agar plate 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, blood plate medium (TSA + 5% defibrinated sheep blood), 18-24h.

Recovery steps:

(1) Prepare 1-2 of above mentioned plates;

(2) Open it in the biosafety cabinet, sterilize of the ampoule, heat the tip in a flame, quickly drop sterile water to creak it, then break it with forceps;

(3) Draw 0.5ml of sterile water into the freeze dried ampoule, make it fully dissolved, and distribute the solution to the plates well in 200ul/plate;

(4) Put the plates under the above culture conditions for cultivation for 18-24hours.

Recovery record: According to the recovery instructions, the results of the recovery are reported as follows:

Item	test results
viability :	good viability， in 24hours strain layer is obvious;colony is typical on the streaked plate
colony morphology: (above)	size: 2-4mm shape: round edge: irregular transparency: opaque color: white bulge: middle bulge surface: bright and smooth texture: moist and easy to stir
Conclusion:	good viability, no abnormal colony morphology, qualified

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