Quick sensitive detection method of potato viruses
A potato virus, sensitive detection technology, applied in the field of plant virology detection, can solve the problems of inability to formulate disease prevention measures, detection sensitivity cannot better determine the infection of different viruses, and affect potato yield and quality
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[0043] 1. Primer design:
[0044] According to the sequences of Potato virus Y, Potato S virus, Potato virus A and Potato leafroll virus in GenBank, select the more conservative regions in a single virus to design primers, optimize the internal pairing and aggregation effects of different primer pairs through experiments between different viruses, and screen Different virus detection primer pairs available for multiplex PCR (see Table 1).
[0045] 2. Coating virus-specific antibodies
[0046] Through testing and optimizing conditions, add polyclonal antibody dilutions of potato virus Y, potato virus S, potato virus A and potato leafroll virus at final concentrations of 1:5000, 1:5000, 1:5000 and 1:5000 respectively in the PCR tube After coating 100 μL at 4°C overnight or at 37°C for 1 h, suck out the liquid in the PCR tube, and treat the PBST solution with DEPC [phosphate buffer containing 0.01% Tween 20 (0.1 mol / L, pH 7. 0) was added to 0.2% DEPC (diethyl pyrocarbonate) and...
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