Construction method of SSR fingerprint suitable for identification of rice hybrid authenticity
A fingerprint, hybrid technology, applied in biochemical equipment and methods, microbial determination/inspection, etc., can solve the problems of simple, fast and accurate identification of the authenticity of hybrids not suitable for crops, and achieve stable results and simple operation. , detect fast effects
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[0026] In this example, the rice hybrid Y Liangyou 689 and its female parent Y58S and male parent R689 seeds were used as test materials, and the seedling genomic DNA was extracted as a template. SSR primer RM211 was used for SSR-PCR amplification, and the SSR-PCR product was high. Resolution melting curve analysis, using GeneScan software for data analysis, and using the constructed SSR fingerprint map for authenticity identification of rice hybrid Y Liangyou 689.
[0027] The specific steps are as follows:
[0028] (1) Genomic DNA extraction
[0029] Using rice hybrids and their parent seeds as materials, young leaves were taken at the seedling stage to extract DNA by CTAB method, and a portion of the DNA solution was taken to measure its concentration, diluted to 10ng / μL, and stored at -20°C for later use.
[0030] (2) SSR-PCR amplification and high-resolution melting curve analysis
[0031] Using 10μL reaction system, containing Mg 2+ (25mM) 1μL, SSR primer RM211 upstream and downs...
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