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A method for identifying Bacillus coagulans

A Bacillus coagulans, a pair of technology, applied in the field of microbial identification, can solve the problems of long detection time, cumbersome biochemical identification operations, etc., to achieve the effect of saving experimental resources

Active Publication Date: 2016-01-06
JIANGSU HFQ BIO TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] To sum up, whether it is used as a probiotic or as the culprit of rancidity, a rapid identification method is needed, and the traditional biochemical identification is cumbersome and takes a long time to meet the needs of various fields. , it is necessary to study the rapid detection of Bacillus coagulans

Method used

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  • A method for identifying Bacillus coagulans
  • A method for identifying Bacillus coagulans
  • A method for identifying Bacillus coagulans

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Experimental procedure

[0035] 1. Extract Bacillus coagulans, Bacillus subtilis, Bacillus cereus, Bacillus megaterium, Bacillus licheniformis, Escherichia coli standard strain (Escherichia coliATCC25922), grape aureus The DNA of Staphylococcus aureus, Pneumococcus and Haemophilus influenzae, the sources of the above strains are shown in Table 1.

[0036] 2. Using the bacterial DNA in step 1 as a template, carry out PCR amplification with HFQ1 / HFQ2 primers.

[0037] The PCR amplification system is: 20 μl by 2 μl 10×Buffer (withMgCl 2 ), 1 μl ldNTP (2.5mmol / L), 1 μl HFQ1 (10mmol / L), 1 μl HFQ2 (10mmol / L), 0.1 μl TaqDNA Polymerase (5U / μl), 1 μl DNA template, 13.9 μl ddH 2 O.

[0038] PCR amplification conditions were: pre-denaturation at 94°C for 5min, denaturation at 94°C for 35s, annealing at 50°C for 40s, extension at 72°C for 40s, 30 cycles, extension at 72°C for 10min, and storage at 4°C.

[0039] 3. The above PCR amplification results were verified by 1% agarose ...

Embodiment 2

[0045] Experimental procedure

[0046] 1. Extract DNA from 2 canned tomato samples that have become rancid and 2 soil samples from Haimen City, Jiangsu Province. The sources of the above materials are shown in Table 2.

[0047] 2. Using the bacterial DNA in step 1 as a template, carry out PCR amplification with HFQ1 / HFQ2 primers.

[0048] The PCR amplification system is: 20 μl by 2 μl 10×Buffer (withMgCl 2 ), 1 μl ldNTP (2.5mmol / L), 1 μl HFQ1 (10mmol / L), 1 μl HFQ2 (10mmol / L), 0.1 μl TaqDNA Polymerase (5U / μl), 1 μl DNA template, 13.9 μl ddH 2 O.

[0049] PCR amplification conditions were: pre-denaturation at 94°C for 5min, denaturation at 94°C for 35s, annealing at 50°C for 40s, extension at 72°C for 40s, 30 cycles, extension at 72°C for 10min, and storage at 4°C.

[0050] 3. The above PCR amplification results were verified by 1% agarose gel electrophoresis.

[0051] Form 2 Material source registration form

[0052] material name

source

Soil A

Baojia...

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PUM

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Abstract

The invention relates to a method for identifying bacillus coagulans, and particularly relates to a novel molecular biology method for identifying bacillus coagulans. By adopting the method, a specific primer HFQ1 / HFQ2 is designed for a specific deoxyribonucleic acid (DNA) sequence of a bacillus coagulans gene, polymerase chain reaction (PCR) amplification detection is carried out on genome DNA of a to-be-detected sample by using the primer, and a bacillus coagulans strain can be identified to the level of seeds. A practical method which is more accurate and easier to operate is provided for identifying the bacillus coagulans, and classification and research work of the bacillus coagulans strain can be facilitated.

Description

technical field [0001] The invention relates to a method for identifying microorganisms, in particular to a method for identifying bacillus coagulans, and belongs to the technical field of molecular biology. Background technique [0002] Bacillus coagulans is rod-shaped, blunt at both ends, Gram-positive bacteria, catalase-positive, terminal spores, without flagella. The optimum growth temperature is 45-50°C, and the optimum pH value is 6.6-7.0. It can decompose sugar to generate L-lactic acid, which is a homolactic acid fermenting bacteria. It is a "generally recognized as safe" bacillus lactic acid bacteria approved by the US FDA. Bacillus coagulans is a rising star in the field of probiotics in recent years. In addition to its health benefits such as lactic acid bacteria and bifidobacteria, it also has high stress resistance such as high temperature resistance, acid resistance and bile salt resistance, and has a strong inhibitory effect. Enteropathogenic capacity. [...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68C12Q1/04C12N15/11C12R1/07
CPCC12Q1/686C12Q2531/113
Inventor 符雷郗洪生施腾鑫陆亚怡
Owner JIANGSU HFQ BIO TECH CO LTD
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