BeNa Culture Collection

  • Bacillus subtilis counting standard substance-BNCC

Bacillus subtilis counting standard substance

  • Price: $400
  • number:BNCC359087
  • Packing:Xilin bottle;(4.0±0.8)× 10 ^ 4CFU/bottle
Essential Information Certificate Related Products
Bacillus subtilis counting standard substance
Culture medium Nutritional gravy medium (English name: 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 mgMnSO4 · H2O to culture Bacillus is beneficial to produce spores.
Subculture procedure 1. According to the arrow sign of Xilin bottle cap, open the strain and dissolved liquid; 2. Pour the bacteria tablets into the dissolved liquid, cover the rubber stopper, shake and dissolve; 3. After being fully dissolved, it can be absorbed in proportion according to the test requirements; 4. Please use up immediately after opening, and do not store it;
Growth conditions 37 ℃, aerobic, 18-24h
Storage conditions -20 ℃, kept away from light
Safety level 1
application Pharmacopoeia test
Sharing mode Public welfare sharing

Reference material of Bacillus subtilis Spores Count

Product number: 359087

Production batch: 20210621

Date of setting value: June 15, 2021

Bacillus subtilis (Bacillus Subtilis) has become one of the most studied and applied strains due to its advantages of good stability, strong resistance, and high resuscitation rate. In the intestinal tract of the target animal, it antagonizes pathogenic microorganisms through biological oxygen capture, and produces a variety of digestive enzymes and nutrients to regulate the health of the digestive tract, enhance the immune function of the animal body, prevent the occurrence of diseases, and achieve the goal of promoting The purpose of animal growth and improving feed conversion rate. In the detection of probiotics, the number of spores is an important indicator to measure the quality of probiotics. This standard material provides a physical reference standard for the detection of spore quantity, and can be used for quantity value transfer, evaluation and control test methods, quality inspection, production process inspection, etc. in laboratories related to microbial testing. The applicable standard GB/T 26428-2010 《Testing of Bacillus subtilis in microbial preparations for feed》, etc.

1.Sample preparation

Select and culture Aspergillus niger (Bacillus subtilis, strain number ATCC 6633) to  produces spores, and dispense the spore suspension into penicillin bottles by pasteur pipette, then make it freeze-dried and sealed.

2. Traceability and value setting method

The reference material is determined by multiple organizations using the spreading method. After 10 times series dilution of the standard substance, the number of colonies on the medium plate is counted, and the total number of colonies is calculated by formula, and the unit of measure is CFU/mL(CFU, colony forming unit Colony-Forming Units). By using quality volume etc measurement instruments that is verified and calibrated, and the quantitative measurement methods which meet the requirements of metrology characteristics, it is ensured that the measurement value of this reference material can be traced back to the basic unit of entity number “1” and the national legal unit of measurement “L”.

3. Characteristic value and uncertainty



standard value (CFU/mL)

Extended uncertainty (CFU/mL,k = 2)

BNCC 359087 Bacillus subtilis spore count standard material 4.0×104 0.8×104

The uncertainty of the setting value is obtained by the combination of uncertainty components introduced by the setting method, balance, pipette, dilution factor, uniformity and stability.

4. Uniformity and stability evaluation

According to the JJF1343 national metrology technical specifications (equivalent ISO guide 35), use Aspergillus niger spore counting and spreading method, and evaluate the uniformity of the sample according to the principle of random sampling. The results of variance analysis show that the uniformity of the sample is good; Using the same measurement method, the stability of the sample is evaluated according to the principle of first density and then sparse. The trend analysis results show that it can be stored stably for a long time at -20 ℃, for 7 days at 4 ℃ and can be shipped in ice bags. The reference material is valid for 12 months from the setting date. We will continue to track and monitor the stability of the standard substance. If changes in the value are found during the validity period, the user will be notified in time.

5. Packaging, storage and usage

1. packaging and preservation: the reference material is packed in 3ml penile bottles and sealed after freeze-drying. It is shipped on dry ice. Upon receipt, check if the dry ice is exhausted or not. Then put it in a refrigerator at -20 ℃.

2. use: the reference material should be used up at one time after opening the bottle. Before use, take it out of -20 ℃ and balance it at room temperature for 10 minutes. accurately weigh (1.09±0.01) g (20°C) sterile distilled water with a balance to reconstitute the lyophilized standard substance, and keep it in a water bath at 80°C for 10 minutes before use.

3. This standard material contains microorganisms, and masks and latex gloves should be worn before use; precautions should be taken to avoid inhalation or the standard value (CFU/mL) and expanded uncertainty (CFU/ mL, k=2) BNCC 359088 Bacillus subtilis spore count standard material 1.5×104 0.3×104 Skin contact; consumables such as remaining samples, packaging bottles and sampling tips after use should be autoclaved as biological pollution waste.

6. References

[1] GB T 26428-2010 Standard for the detection of Bacillus subtilis in forage microbial preparations [S].


1. The reference material is only for laboratory research and analysis and testing. Complaints arising from improper use or storage by users shall not be liable.

2. Please check the variety, quantity, packaging and certificate immediately upon receipt. The relevant compensation is limited to the reference material itself and does not involve any other losses.

3. The reference material is shipped with the certificate stamped”"special seal for certified reference materials of Beijing Beina Chuanglian Biotechnology Research Institute". We will not accept any complaints if the certificate can not be provided. Please keep this certificate properly.

4. For more application-related information, please contact the technical department.

Beijing Beina Chuanglian Biotechnology Research Institute

Technicians: 400-6699-833

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