SNP marker related with millet tiller number character and detecting primer and application thereof
A tiller number and marker technology, which is applied in the determination/inspection of microorganisms, biochemical equipment and methods, recombinant DNA technology, etc., and can solve the problems that the development and application of SNP molecular markers have not been reported.
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Embodiment 1
[0033] Example 1 High-throughput sequencing and SNP marker information analysis
[0034] In this example, the material of the male parent "Jizhanggu No. 1" and the female parent "A2" of millet, the F1 generation material of the cross between the two, and the 441 materials of the F2 population of 13 generations of self-inbred, that is, the recombinant inbred lines (Recombinant inbred lines) , abbreviated RILs). All materials were planted in Zhangjiakou, Hebei Province, and the data of tiller numbers and other traits were recorded and sorted. After degenerate genome resequencing of each material, the alignment of the reference genome was completed, and SNP markers were obtained. By analyzing the tiller number trait data, the SNP markers related to it 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 individual sample, a total of 444 samples, including 441 RILs, 2 parents...
Embodiment 2
[0058] Example 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 300bp before and after the SNP site, design and develop SNP marker primers, and perform PCR amplification with the DNA of the male and female parent materials as templates. The amplification products with normal primer amplification and the predicted size of PCR amplification products were selected for gel cutting recovery and sequencing, and labeled primers with SNP site differences in the amplified gene sequences of the male and female parent materials were selected. Using the F1 and F2 generation materials as templates, PCR amplification of SNP marker primers and sequencing results were performed, and marker primers that were consistent with genotyping and related to phenotypic traits were selected.
[0060] First, according to the sequencing results after the recovery of the PCR p...
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