Multi-PCR detection kit capable of simultaneously detecting clostridium perfringens, clostridium hemolyticumbovis and B-type clostridium novyi and application of multi-PCR detection kit
A technology of Clostridium perfringens and Clostridium novyi, which is applied in the determination/testing of microorganisms, biochemical equipment and methods, etc., and can solve the problem of false positives and the inability to simultaneously detect Clostridium hemolyticus and Noviella type B Clostridium, expensive reagents and other issues, to achieve good specificity and sensitivity
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[0030] 1. Primer design: Download the α-toxin gene of Clostridium perfringens (accession number: L43548) from the website http: / / www.ncbi.nlm.nih.gov / genbank / , use this gene as a template, and use Primer5.0 The software designed a pair of primers: PrimerF1: 5´-GCTAATGTTACTGCCGTTGA-3´, PrimerR1: 5´-CCTCTGATACATCGTGTAAG-3´.
[0031] According to Clostridium hemolyticus flagellin gene FliA(C), FliA(H) (gene accession number: AB058931, AB058939), Clostridium novyi type B flagellin gene FliA (gene accession number: AB058938) designed 3 primers: PrimerF2: 5´-AGAATAAACAGAGCTGGAGATG-3´; PrimerR2: 5´-TTATGCTAACTTTAGCTG CGTC-3´; PrimerR3: 5´-CTGCTGTACCTTCTATGAACC-3´.
[0032]2. Kit assembly: use the primer design software (Primer 5.0) to design the required 5 primers, and then send them to the biological company for chemical synthesis. Dilute the synthesized 5 primers to 10 µM with sterile water and mark them as F1, F2, F3, F4 and F5; purchase 1 bottle (500 µL) of commercial GoTaq DNA ...
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