Bacillus subtilis subsp.|BNCC353544 |BNCC

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Bacillus subtilis subsp.

Culture medium	Brain heart extract medium (BHIA/BHIB): brain extract 5.0g, heart extract 12.5g, peptone 5.0g, tryptone 10.0g, sodium chloride 5.0g, glucose 2.0g, disodium hydrogen phosphate 2.5g, agar 20.0g (without liquid medium),pH 7.4±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°C, 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

No. : 353544 / ATCC 6633

Product format: quantitative bacteria tablets

Number of live bacteria : 100 ~ 500CFU/bottle batch: 190916

Valid until : 2020.9.15

Date of inspection : 2019.9.16

Sampling ratio : ≥2%

Report number : RS19353544-2

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

Item	description	test results
staining microscopy	gram-negative and positive, cell morphology observed under microscope	G + bacillus, in the shape of single rod, double rod and connecting rod
colony morphology	observation of single colony morphology	light pink, rough, low flat, irregular edges
purity	non-selective medium culture, colony observation and microscopic examination	No bacteria
Reference value of live bacteria	non-selective medium plate, live bacteria count	360CFU/bottle
phenotype and genotype analysis	phenotypic testing, gene sequencing	qualified
test conclusion	after quality inspection by BNCC quality control testing system, the product is qualified

Product list :1 bottle of 100 ~ 500CFU quantitative strain, 1 bottle of 1mL dissolved solution and 1 copy of strain certificate;

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

Handling procedure:

1 Open the penicillin-bottle according to the arrow indication on the vial cap;
2 Transfer the strain tablets into the vial of solution, capped with rubber plug, and shake to dissolve
3 It will be 100-500cfu/ml after full dissolved, and use appropriate amount according to the operation demand
4 The bacterial tablets shall be used up once and shall not be retained once the penicillin bottled is opened.

Notes:

1 This product is a quantitative strain. Improper storage conditions and handling procedure will affect the accuracy of the strain quantities. Therefore, please operate in strict accordance with the above mentioned instructions.
2 Waste generated from the handling process should be discarded after high-pressure sterilization