A method for identifying the purity of Jingke 968 corn hybrid based on SNP markers
A corn and molecular marker technology, applied in the field of crop molecular biology, can solve the problems of heavy workload, difficult high-throughput detection methods, and long identification period
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Embodiment 1
[0037] Example 1 Determination of SNP molecular markers and primers for identifying the purity of Jingke 968 corn hybrids
[0038] (1) Basic site: Using the 384 SNP sites published in the "Maize DNA Fingerprint Library and Variety Molecular Identification SNP Core Site Combination-maizeSNP384" (published in Chinese Patent 201410756086.0) as the starting site set, and established SNP-DNA fingerprint data of 335 maize hybrids under national review.
[0039] (2) SNP site test and biological characteristics evaluation: 192 corn hybrids were selected, and DNA was extracted by rapid method, CTAB, kit, and various methods. Based on the KASP technology system, the DNA extracted by different methods was used to verify the site. The typing effect, repeatability, and stability of the points are used to obtain a set of candidate sites.
[0040] (3) Evaluation of SNP loci polymorphism: using data of 200 different types of maize inbred lines and 335 maize hybrids, the polymorphism paramete...
Embodiment 2
[0049] Embodiment 2 utilizes the SNP molecular marker provided by the present invention to identify the method for the purity of Jingke 968 corn hybrid
[0050] Using the primer combinations designed in Example 1 for the 4 SNP sites, the scheme for identifying the seed purity of Jingke 968 is as follows: Synthesize primers according to the KASP technical requirements, and the primers are common primers without fluorescent groups; PCR amplification system MasterMix; configure the reaction system to add DNA, primers and MasterMix; run the amplification reaction program; scan the fluorescence signal in situ; perform data analysis to obtain genotype data; The genotype data is used to determine whether the individual plant is a normal plant, an inbred seedling, or an allotype plant, and finally its purity value is comprehensively determined. The specific operations are as follows:
[0051] (1) Preparation of the sample to be tested:
[0052] At least 150 seeds are randomly select...
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