Klebsiella pneumoniae|361007 |BNCC

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Klebsiella pneumoniae

Subculture procedure	① Prepare one test tube containing 5-10 mL of liquid culture medium and two plates; ② Open the safety cabinet, burn the top with an alcohol lamp, quickly drip sterile water to break it, and then use tweezers to crush it; ③ Transfer 0.5mL of liquid culture medium into a freeze-drying tube, dissolve it thoroughly, and then transfer it back into the liquid test tube, mixing well; ④ Transfer 0.2mL of bacterial suspension into a Agar plate, apply evenly, and repeat twice to obtain two plates; ⑤ Place all liquid test tubes and plates under the above cultivation conditions, and the bacterial strains can be used once they grow.
Growth conditions	37 ℃; 18-24h; aerobic;
Storage conditions	2-8 ℃
Safety level	2
morphology	The colony diameter is 1-2mm, circular, with neat edges, opaque, gray white on the front, raised in the middle, bright on the surface, easy to pick up, G - (red), rod-shaped
Sharing mode	Public welfare sharing

Klebsiella pneumoniae

Storage conditions: 2~8 ℃

No. 361007

Product format： freeze dried, 200ul

Validity period: 6 years

Biosafety level: 1, 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 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, tryptone soybean medium, 18-24h. Tryptone soybean medium: tryptone 17.0g, soy 3.0g, sodium chloride 5.0g, dipotassium hydrogen phosphate 2.5g, glucose 2.5g, agar 20.0g (not included in liquid medium), distilled water 1.0L, pH 7.3 ±0.2. Sterilize at 121℃ for 15min.

Recovery steps:

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

(2)Open the ampoule 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 24 hours liquid medium become turbid, obvious strain layer occurs on the plate; colony is typical on marked plate
colony morphology:	Small: 2-4mm Shape: Round Edge: Neat Transparency: Opaque Color: white bulge: middle bulge surface: bright and smooth texture: moist and viscous
Conclusion:	good viability, no abnormal colony morphology, qualified;

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