SNP marker relevant to stem diameter character of millet, detection primers for SNP marker, and applications
A technology of labeling and millet, which is applied in the determination/inspection of microorganisms, biochemical equipment and methods, recombinant DNA technology, etc., and can solve the problems of unreported research on the development and application of SNP molecular markers.
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Embodiment 1
[0033] Example 1 High-throughput sequencing and SNP marker information analysis
[0034] In this example, the materials of the male parent "Ji Zhanggu No. 1" and the female parent "A2" of millet, the F1 generation materials of the two hybrids, and 441 materials of the F2 population of the 13th generation of selfing, that is, recombinant inbred lines (Recombinant inbred lines , abbreviated as RILs). Each material was planted in Zhangjiakou, Hebei, and the trait data such as stem diameter were recorded and sorted out. After the degenerate genome resequencing of each material, the alignment of the reference genome was completed, and the SNP marker was obtained. By analyzing the data of stem thickness traits, the SNP markers related to them were obtained, and the SNP markers were verified by clone sequencing.
[0035] In this example, the RADseq method was used to extract the genomic DNA of each sample individual, a total of 444 samples, including 441 RILs, 2 parents, and 1 F1 i...
Embodiment 2
[0058] Embodiment 2 SNP marker verification
[0059] According to the predicted bin marker and SNP site, compare the reference genome to obtain the gene sequence of the relevant region, select about 300 bp before and after the SNP site, design and develop SNP marker primers, and use the DNA of the paternal and maternal materials as templates for PCR amplification. Select the amplified products with normal primer amplification and the compounded predicted size of the PCR amplification products for gel cutting recovery and sequencing, and select the labeled primers with SNP site differences in the amplified gene sequences of the paternal and maternal materials. Using the F1 and F2 generation materials as templates, carry out PCR amplification of SNP marker primers and sequencing of the results, and select marker primers that are consistent with genotyping and phenotypic traits.
[0060] First, according to the sequencing results after PCR product recovery, the markers for the di...
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