Denitrifying subspecies of colorless bacillus xylose oxidizing|BNCC221600 |BNCC

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Achromobacter xylosoxidans subsp. denitrificans

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 flask containing LB liquid medium; (2) Open the ampoule 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 medium, mix evenly; cultivate in a shaker at 30 ℃ (140r /min) for 18-24hours.
Growth conditions	30 ℃,18-24h, aerobic
Storage conditions	2-8 ℃
morphology	Size: general color: yellowish shape: round edge: neat edge Wet and Dry: Wet Smooth: Smooth Transparency: Opaque Protuberance: Protuberance
Separation substrate	Oil contaminated soil
application	Growth with phenanthrene, anthracene and fluorene as sole carbon sources
Sharing mode	Public welfare sharing

Achromobacter xylosoxidans subsp.

BNCC No.：221600

Storage conditions: 2~8 ℃

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. 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:30 ℃, aerobic, nutritious gravy medium (NB). Nutritional gravy medium: beef paste 3.0g, peptone 10.0g,NaCl 5.0g, distilled water 1.0L, pH7.0. 121 ℃,15min.

Recovery steps:

(1)Prepare a flask of NB liquid media

(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 30℃ for 18-24 hours (140r/min);

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

item	test results
viability	good viability,in 20h,NB bacterial fluid turbidity; The bacterial liquid is marked NA plate with obvious colony
colony morphology:	size: general color: yellowish shape: round edge: neat edge wet and dry: wet and smooth: smooth transparency: opaque uplift: bulge
conclusion:	good viability, and colony morphology is not abnormal, completely consistent with the above figure, qualified.

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