Multiple fluorescent quantitative PCR detection primer and reagent kit for novel goose astrovirus, goose paramyxovirus and goose parvovirus, and application
A multiplex fluorescence quantitative, goose paramyxovirus technology, applied in the field of molecular biology, to achieve high feasibility and application prospects, good reproducibility, high specificity and specificity
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[0050] 1.1 Primer design and synthesis
[0051] The sequence refers to the sequence of each target gene in the Gene Bank database, and 3 sets of specific primers and probes that can amplify 3 goose pathogens are designed. The primers and probes are designed by ABI's Primer Express Software v2.0 and are designed by BGI Synthesized by the company, store at -20℃ for later use.
[0052] The nucleotide sequences of the three sets of primers and probes are shown in Table 2 below:
[0053] Table 2 Nucleotide sequences of primers and probes
[0054]
[0055] 1.2 Viral nucleic acid extraction and cDNA synthesis
[0056] Use Easy Viral DNA / RNA Kit (spin column type viral DNA / RNA extraction kit) to extract three viral DNA / RNA, and use HiScript cDNA first-strand synthesis kit to reverse transcription of three viral RNAs: 40μL system , Add 16μL of RNA template, 2μL of random primers, 2×RTMIX 20μL, HiScript EnzymeMix 2μL to the EP tube, total volume 40μL, and place in PCR machine for cDNA synthesis...
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