Real-time fluorescence PCR detection method and kit for sudden death syndrome virus of soybean
A detection kit, soybean sudden death technology, applied in the direction of fluorescence/phosphorescence, biochemical equipment and methods, microbial measurement/inspection, etc., to achieve the effect of short detection time, high sensitivity and strong specificity
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Embodiment 1
[0060] 1. Experimental reagents
[0061] Universal Real-time PCR Master Mix was purchased from ABI Company, and DNA extraction kit DNeasy Plant Mini Kit was purchased from QIAGEN Company.
[0062] Primers and probes were synthesized by Shanghai Chaoshi Biotechnology Co., Ltd.
[0063] 2. Experimental equipment
[0064] ABI 7700 real-time fluorescent PCR instrument.
[0065] 3. Experimental method and process
[0066] (1) Design of primers and probes
[0067] First, compare and analyze the genome sequence information of the North American soybean sudden death syndrome pathogen and the South American soybean sudden death syndrome pathogen and their similar species and related species to find out the unique and stable specific base sites of each detection target sequence , the designed alternative primers and probes are shown in Table 1;
[0068] Table 1 Primers and probes for testing
[0069] Numbering
Sequence (5'→3')
Seq ID NO.
FV-F
5′-GGTGCA...
Embodiment 2
[0094] A detection kit is used for real-time fluorescent PCR detection of North American soybean sudden death syndrome pathogen. The kit contains the following primers and probes:
[0095] Primer
[0096] probe
[0097] The kit also includes instructions for use.
Embodiment 3
[0099] A detection kit is used for real-time fluorescent PCR detection of South American soybean sudden death syndrome pathogen. The kit contains the following primers and probes:
[0100] Primer
[0101] The kit also includes instructions for use.
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