A kind of HDA primer of Clostridium perfringens in food and application method thereof
A Clostridium perfringens, food technology, applied in the direction of microorganism-based methods, biochemical equipment and methods, microorganisms, etc., can solve the problem that nucleic acid amplification technology cannot be more widely promoted and applied, and achieve considerable economic Benefits, saving manpower and material resources, and the effect of rapid detection methods
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Embodiment 1
[0031] Example 1: Design of C. perfringens HDA primers
[0032] The primers were synthesized by Bao Bioengineering (Dalian) Co., Ltd. The sequences of the two primers are shown in Table 1.
[0033] Table 1 Clostridium perfringens HDA primer sequence list
[0034] Primername
Embodiment 2
[0035] Example 2: Application of primers in LAMP detection of Staphylococcus aureus
[0036] (1) Extraction of genomic DNA from Clostridium perfringens
[0037] Pick out Clostridium perfringens and add it to a conical flask with 1% peptone water, shake it slowly by hand until it is fully mixed, and incubate at 36℃±1℃ for 24h±2h. Pipette 1mL of transfection into 10mL SPS, and incubate at 6℃±1℃ for 24h±2h. Finally, 1 mL of the enrichment solution was taken to extract the genomic DNA of Clostridium perfringens with a bacterial genomic DNA extraction kit, and stored at -20°C for later use.
[0038] (2) Optimization and establishment of HDA reaction system and reaction conditions
[0039] A one-step HDA reaction system was used. The reaction system for detecting Clostridium perfringens by HDA method was: 5 μL of 10×buffer (100 mM dithiothreitol, 350 mM Tris-HAc, 100 mM MgSO) 4 , 1mg / mL BSA), 0.04μmol dNTPs, 0.16μmol ATP, 10U Bst ploymerase, 0.10~0.30μg UvrD helicase, 1.0~5.0μg ...
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