PCR primer, PCR detection kit and detection method for detecting B.xylophilus
A technology of pine wood nematode and detection method, applied in biochemical equipment and methods, measurement/testing of microorganisms, DNA/RNA fragments, etc., capable of solving expensive, high-cost, radioactive and other problems
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Embodiment 1
[0029] Embodiment 1 detects the design of the PCR primer of pine xylophilus, screening and the establishment of detection method
[0030] 1. Materials and methods
[0031] 1.1 Test material
[0032] 1.1.1 Biomaterials
[0033] Monochamus monochamus and Spruce monochamus collected in the field from 2017 to 2019 (Table 1); B. xylophilus and B. xylophilus cultivated in the laboratory (Table 2).
[0034] Table 1 Monochamus monochamus and Spruce monochamus collected in the field
[0035]
[0036] Table 2 B. xylophilus and B. xylophilus cultivated in the laboratory
[0037]
[0038]
[0039] 1.1.2 Chemical reagents
[0040] Liquid nitrogen; TE buffer solution; PrimeSTAR HS DNA (Premix) (TaKaRa).
[0041] 1.2PCR amplification
[0042] 1.2.1 PCR primer design
[0043] Using the Bx-cpi gene as the target gene, primers specific to B. xylophilus were designed by Primer 6.0. 1.2.2 Establishment of PCR reaction system
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