Escherichia coli|133721 |BNCC

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Escherichia coli

Subculture procedure	① Prepare 2 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 Suck 0.5mL sterile water into the freeze-drying tube, dissolve thoroughly, and mix well; ④ Transfer 0.2mL of bacterial suspension into a Agar plate, apply evenly, and repeat twice to obtain two plates; ⑤ Place all the plates under the above cultivation conditions, and once the strains grow, they can be used.
Growth conditions	37 ℃; 18-24h; aerobic
Storage conditions	2-8 ℃
Safety level	1
morphology	The colony diameter is 1-2 mm, circular with regular edges and opaque. The obverse side is creamy white, while the reverse side is yellow, centrally raised with a smooth surface and moist texture, easily picked off. No pigment production is observed. Gram-negative (red), rod-shaped bacteria, purity: pure.
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Escherichia coli

Storage conditions : 2~8 ℃

No. : 133721

Product format : freeze dried,200ul

Validity period : freeze-dried tube 24 months

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. 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.

Recovery/subculture:37°C, aerobic, nutrient gravy medium. 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 24-48 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 become turbid, bacterial fluid scribing plate colonies are obviously typical
colony morphology	size: general color: off-white shape: round edge: neat edge wet and dry: wet and smooth: smooth transparency: opaque uplift: uplift
Conclusion	good viability, and colony morphology is not abnormal, completely consistent with the above figure, qualified

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