Escherichia coli EHEC|BNCC345676 |BNCC

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Escherichia coli EHEC

Culture medium	Nutritional gravy medium (NA/NB): beef paste 3.0g, peptone 10.0g,NaCl 5.0g, agar 20.0g (without liquid medium), distilled water 1.0L,pH 7.0. Sterilization at 121 ℃ for 15min. [Note] Adding 5 mg MnSO4 · H2O to culture Bacillus is beneficial to produce spores.
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;
Growth characteristics	Gram-negative bacilli. Protein secretion regulatory gene (escV), Shiga toxin I gene (stx1 or stx1A) and Shiga toxin II gene (stx2 or stx2A).
Storage conditions	2-8 ℃
morphology	The colony diameter is 1-2mm, round, with neat edges, yellow on the front, light color, yellow on the back, light color, flat shape, bright surface, smooth surface, easy to pick up, G-(red)
application	"GB/T 4789.6-2016 Diarrhea Escherichia coli Test" Positive Control Strain., "GB 4789 National Food Safety Standard Food Microbiological Inspection" Standard Strain Recommendation
Sharing mode	Public welfare sharing

Escherichia coli EHEC

Storage conditions : 2~8 ℃

No. : 345676

Product format: freeze dried, 200ul

Validity period: 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 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 lose of viability.

Growth conditions :37 ℃, aerobic, nutrient gravy medium, 18-24h. Nutritional gravy 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. Sterilization at 121 ℃ for 15min.

Recovery steps:
(1)Prepare a test tube containing 5-10mL of liquid culture medium and 2 plates;
(2)Open 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: 2-4mm shape: round edge: neat transparency: opaque color: light yellow uplift: flat surface: bright and smooth texture: moist and easy to stir
conclusion	good viability, no abnormal colony morphology, qualified

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