Bacillus pseudoharens|BNCC186420 |BNCC

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Bacillus pseudoharens

Culture medium	Alkaline nutrient agar medium (A-NA): alkaline nutrient agar medium: beef paste 3.0g, peptone 10.0g,NaCl 5.0g, agar 20.0g, distilled water 1.0L,pH 7.0. After sterilization at 121 ℃ for 15min, the culture medium was adjusted to pH9.7 with 1mol/L sodium hemicarbonate sterile solution (formula attached). [Note] Sodium sesquicarbonate solution: NaHCO3 4.2g, anhydrous Na2CO3 5.3g, distilled water 0.1L.
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	Size: 1-2mm Shape: Round Edge: Neat Transparency: Opaque Color: light yellow uplift: flat surface: smooth texture: wet
Separation substrate	Soil on the shore of the lake
Sharing mode	Public welfare sharing

Bacillus pseudofirmus

Storage conditions: 2~8 ℃

No. 186420

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 plate 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, alkaline nutrient agar medium, 18-24h. Alkaline nutrient agar medium: After sterilizing the nutrient agar, adjust the medium to pH 9.7 with 1 mol/L sodium sesquicarbonate sterile solution (recipe attached). [Note] Sodium sesquicarbonate solution: NaHCO3 4.2g, anhydrous Na2CO3 5.3g, distilled water 0.1L.

Recovery steps:

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

(2) Open the ampoule 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 recovery are reported as follows:

Item	test results
viability:	good viability, in 18-24hours liquid medium become turbid,strain layer is obvious;colony is typical on the streaked plate
colony morphology: (above)	size: 1-2mm ; Shape: round ; Edge: neat ; Transparency: opaque color: light yellow ; Protuberance: flat ; Surface: smooth ; Texture: moist
Conclusion:	good viability,no abnormal colony morphology, qualified