Gene chip for detecting screening-gene-Hpt-containing transgenic plant and product and application thereof
A technology for genetically modified plants and screening genes, which is used in the determination/inspection of microorganisms, biochemical equipment and methods, fluorescence/phosphorescence, etc. It can solve the problems of patent reports on gene chip detection technology that have not yet discovered the Hpt gene, and achieve good specificity. Sensitivity, high sensitivity, and the effect of improving the level of supervision
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[0039] 1. Primer and probe design
[0040] According to the Hpt gene sequence, the Primer Premier 5.0 primer design software was used to design the Hpt-specific detection primers for the screening gene, and the primers were screened and optimized. The fluorescently labeled primers are fluorescently labeled with Cy3-Amidite at the 5' end. The primers were synthesized by Shanghai Boya Biological Company, and the primers were diluted to 10 μmol / L for use.
[0041] Use Primer premier 5.0 and Oligo 6.0 software to design 40mer Oligo candidate probes based on the sequence of Hpt. The general principles of design are: (1) have relatively similar TM values, and the fluctuation range is 5°C; (2) GC content is 45% ~55%, otherwise the background of non-specific hybridization will be increased; (3) the repeated single bases shall not exceed 4 consecutively; (4) the length of the paired bases of the most stable secondary structure of the probe molecule shall not be greater than 4bp. The ...
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