Enterotoxigenic Escherichia coli|186734 |BNCC

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Escherichia coli ETEC

Subculture procedure	① Prepare 1 tube containing 5–10 mL liquid medium and 2 plates; ② Open in a safety cabinet, cauterize the top with an alcohol lamp, then quickly add sterile water to shatter it; break into pieces with forceps; ③ Pipette 0.5 mL liquid medium into the freeze-dried vial, dissolve completely, then return to the liquid tube and mix thoroughly; ④ Pipette 0.2 mL of the bacterial suspension onto a Agar plate and spread evenly. Repeat twice to obtain two plates. ⑤ Incubate all liquid tubes and plates under the specified conditions. The culture is ready for use once bacterial growth appears.
Growth conditions	37°C; 18-24h; aerobic;
Storage conditions	2-8°C
Safety level	2
morphology	Single colonies measure 1–2 mm in diameter, are circular in shape with neat, opaque edges, grayish-white in color, and exhibit a raised center.
Sharing mode	Public welfare sharing

Escherichia coli ETEC

Storage conditions : 2~8 ℃

No. : 186734

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.

Culture conditions :37°C, aerobic, nutrient agar medium, 18-24h. 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 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 cell recovery are reported as follows:

item	test result
viability	good viability，in 20 hours liquid medium become turbid, obvious strain layer occurs on the plate,colony is typical on marked plate.
colony morphology: (above)	size: 2-4mm shape: round edge: neat transparency: translucent color: light yellow uplift: middle convex surface: gray texture: viscous
conclusion	good viability，no abnormal colony morphology,qualified

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