Quantitative strain of Bacillus subtilis|BNCC361421 |BNCC

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Quantitative strain of Bacillus subtilis

Culture medium	Tryptone soybean medium (English name: TSA/TSB): tryptone 17.0g, soybean peptone 3.0g, sodium chloride 5.0g, dipotassium hydrogen phosphate 2.5g, glucose 2.5g, agar 20.0g (without liquid medium), distilled water 1.0L,pH 7.3±0.2. Sterilization at 121 ℃ for 15min.
Subculture procedure	(1) According to the culture conditions, the bacteria are first cultured into a bacterial suspension; ② Measure the absorbance of the bacterial suspension and calculate the initial concentration of the bacterial suspension according to the standard formula; (3) The initial concentration multiplied by the survival rate is the actual concentration after freeze-drying; (4) Dilute the bacterial suspension according to the actual concentration after freeze-drying to obtain the target concentration, and then subpack and freeze-dry;
Growth conditions	30 ℃;18-24h; Aerobic;
Storage conditions	-20 ℃
morphology	The colony diameter is 1-2mm, round, with neat edges, opaque, gray-white on the front, gray-white on the back, convex in the middle, bright surface, smooth surface, moist texture, easy to stir up, G + (blue-purple), bacilli, purity: pure
Sharing mode	Public welfare sharing

Bacillus subtilis subsp.

Strain number: BNCC 361421 /ATCC6633

Product format: quantitative bacteria tablets

Number of viable bacteria:(4.7±0.3)×10⁷CFU/bottle

Product batch: 220428

Valid until: 2023.04.27

Date of inspection: 2022.04.27

Sampling ratio:≥2%

Report number: RS22293727-1

Biosafety level: 1, handle in ultra-clear table or safety cabinet

items	description	results
stain microscopy	Gram-negative, positive, cell morphology observed under microscope	G + cocci
colony morphology	Observe the morphology of single colonies	golden yellow, raised, bright, moist
purity	Non-selective culture medium, colony observation and microscopy	No contaminated bacterial
Reference value of live bacteria	Non-selective media plates, viable count	(4.7±0.3)×10⁷CFU
Phenotypic and Genotypic Analysis	Phenotypic testing, gene sequencing	qualified
test conclusion	After the BNCC quality control inspection system, the product is qualified

Product list: quantitative strains 1 bottle, 1mL lysate 1 bottle, 1 strain certificate;

Validity period: unopened, below -20 ℃, 12 months.

Usage:

1. Open the strain and lysate according to the arrow indication of the vial cap;

2. Pour the bacterial flakes into the dissolving solution, cover with a rubber stopper, and shake to dissolve;

3. After fully dissolved, it is (4.7±0.3)×10⁷CFU/mL, and it can be drawn in proportion according to the test requirements;

4. Please use up immediately after opening and do not store;

Notes:

① This product is a quantitative strain. Improper storage conditions and use methods will affect the accuracy of the number of strains, so please operate in strict accordance with the above storage conditions and methods.

②Used germ-carrying consumables should be discarded after autoclaving.