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Kit extraction method suitable for Staphylococcus aureus genomes

A staphylococcus, extraction method technology, applied in the biological field, can solve the problems of biosafety, lack of reference materials for food microorganisms, etc., and achieve the effect of improving concentration and purity

Inactive Publication Date: 2020-05-26
INST OF AGRO FOOD SCI & TECH SHANDONG ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, food microbiological reference materials are generally scarce in the world. At the same time, due to biosafety issues, there are many difficulties in the international sharing of these reference materials.

Method used

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  • Kit extraction method suitable for Staphylococcus aureus genomes
  • Kit extraction method suitable for Staphylococcus aureus genomes
  • Kit extraction method suitable for Staphylococcus aureus genomes

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] 1.1 Experimental strains and characteristic virulence gene primers are shown in Table 1. For Salmonella Enteritidis and Staphylococcus aureus virulence gene primers, refer to the industry standard SN / T 1869-2007.

[0036] Medium: Beijing Luqiao BHI broth medium.

[0037] Agarose: Beijing Quanshijin Company.

[0038] RNase A: Tiangen RT405-12.

[0039] Bacterial Genomic DNA Extraction Kit: OMEGA Bacterial Genomic DNA Extraction Kit (D3350-01); Tiangen Bacterial Genomic DNA Extraction Kit (DP302).

[0040] PCR reaction reagent: 2×Taq PCR premix reagent II (KT211) (Tiangen Biochemical Technology (Beijing) Co., Ltd.).

[0041] Bacterial strains and primer sequences used in the present invention of table 1

[0042]

[0043] 1.2 The extraction method before improvement is as follows:

[0044]After centrifuging and discarding the supernatant of 1 mL of the bacterial suspension cultured overnight, add lysozyme solution to break the wall, the specific method is: add 110 ...

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Abstract

The invention belongs to the technical field of biology, and particularly relates to a kit extraction method suitable for Staphylococcus aureus genomes. The kit extraction method suitable for Staphylococcus aureus genomes is characterized in that Staphylococcus aureus is washed with normal saline, thalli are subjected to boiling water bath treatment in the normal saline, then lysozyme is adopted for thalli cracking treatment, the lysozyme treatment time is longer than or equal to 12 h, and then the genomes are extracted with a Tiangen kit. When the method is used for extracting the Staphylococcus aureus genomes, the wall breaking effect of Staphylococcus aureus can be remarkably improved, and the purpose of improving the genome concentration is achieved. Through the method, the Staphylococcus aureus genome concentration is obviously improved, the values of A260 / A280 and A260 / A230 reach 1.8-2.0, and the requirements of standard substances are met.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a kit extraction method suitable for the genome of Staphylococcus aureus. Background technique [0002] Foodborne pathogens are pathogenic bacteria that can cause food poisoning or use food as the medium of transmission, which threaten human health and cause serious food safety problems. There are many types of food pathogens, the common ones are Escherichia coli, Staphylococcus aureus, Salmonella and Listeria monocytogenes. The establishment of accurate, sensitive, rapid and specific detection methods for the detection of food pathogens plays an important role in the prevention and control of food microbial food poisoning incidents. [0003] Among the above germs, Staphylococcus aureus is also an important pathogen causing food poisoning, and it is also a worldwide health problem. At present, the disease outbreak mode caused by microbial contamination has also changed ...

Claims

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Application Information

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IPC IPC(8): C12N15/10
CPCC12N15/1003
Inventor 杨金玉陈蕾蕾陈相艳杨焕蝶王易芬刘孝永王军华张彦昊
Owner INST OF AGRO FOOD SCI & TECH SHANDONG ACAD OF AGRI SCI
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