Quantitative strain of Bacillus subtilis|BNCC353782 |BNCC

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Bacillus subtilis

Culture medium	Nutritional gravy medium (NA/NB): beef paste 3.0g, peptone 10.0g,NaCl 5.0g, agar 20.0g (without liquid medium), distilled water 1.0L,pH 7.0. Sterilization at 121 ℃ for 15min. [Note] Adding 5 mg MnSO4 · H2O to culture Bacillus is beneficial to produce spores.
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	37 ℃, aerobic
Growth characteristics	Gram staining is positive, rod-shaped, rarely strands, uniform staining; flagella are lateral and movable; endospores are 0.8 × 1.5-1.8 μm, and the surface of free spores is poorly colored; the colonies on the agar culture medium are round or Irregular shape; the surface color is dark, opaque, can wrinkle, can change to cream or brown; the shape of the colony varies greatly with the composition of the culture medium; strictly nourishing or facultative anaerobic.
Storage conditions	-20 ℃
morphology	gray, flat, dry, G + bacilli
application	Research; Analytical testing. Antibiotic titer determination, temporarily used for the verification of midecamycin. Antibody analysis, positive control strain of GB 4789.2-2010 colony count determination, standard strain of GB 4789.28 medium and reagent quality requirements, quality control strain of Chinese pharmacopoeia, 2015 edition.
Sharing mode	Public welfare sharing

Bacillus subtilis

No. :353782 / CMCC63501

Product format: quantitative bacteria tablets

Number of live bacteria : 100 ~ 500CFU/bottle

Product batch : 191212

Valid until : 2020.6.14

Date of inspection : 2019.12.15

Sampling ratio : ≥2%

Report number: RS19353782-1

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

item	description	test result
stain microscopy	Gram-positive, observe cell morphology under microscope	G + cocci
colony morphology	observation of single colony morphology	Gray, dry, 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	260CFU/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 ℃, 6 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.

Attentions:

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

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