Lysobacter enzymogenes subsp. enzymogenes|BNCC223409 |BNCC

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Lysobacter enzymogenes subsp. enzymogenes

Culture medium	Nutritional gravy medium (NA/NB): beef paste 3.0g, peptone 10.0g,NaCl 5.0g, agar 20.0g (not included in liquid medium), distilled water 1.0L,pH 7.0. Sterilization at 121 ℃ for 15min. [Note] Adding 5 mg MnSO4 · H2O when culturing Bacillus. it 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 safety 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 ℃,18-24h, aerobic
Storage conditions	2-8 ℃
morphology	The colony diameter is 1-2mm, opaque, yellow on the front, convex in the middle, bright surface, smooth surface, moist texture, easy to stir up, G-(red), bacilli
Separation substrate	Soil
Sharing mode	Public welfare sharing

Lysobacter enzymogenes subsp.

No.: 223409

Storage conditions: 2~8 ℃

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 receiving goods, please contact customer service within 24 hours. if it is overdue, it will be regarded as good receiving goods. Dry powder shall not be retained after one use, and can be used after 1~2 generations of activation. please refer to the attached page for details of inclined plane and bacterial solution. Please activate in strict accordance with this instruction, otherwise the replacement service will not be provided if the strain is abnormal and inactivated.

Growth conditions:30°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:

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

②Open it in the safety cabinet, burn the top with an alcohol lamp, quickly drop sterile water to break it, and then break it with tweezers;

③Suck 0.5mL of liquid culture medium into lyophilized tube, fully dissolve and return to liquid test tube again, and mix well;

④Absorb 0.2mL of bacterial suspension into the plate, coat evenly, repeat twice to obtain two plates;

⑤Place all the liquid test tubes and plates under the above culture conditions, and the strains can be used when they grow out.

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

item	test result
viability	good viability,in 20h liquid medium become turbid, obvious strain layer occurs on the plate,colony is typical on marked plate.
colony morphology: (above)	size: 1-2mm shape: round edge: neat transparency: opaque color: yellow uplift: middle raised surface: smooth and bright texture: wet and easy to stir
conclusion:	good viability, no abnormal colony morphology, qualified

inspector/date: boffei 2019.11.28 auditor signature: Sudan

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