SNP molecular marker for identifying Shanghai colorful waxy corn NO.1 and identifying method thereof
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A technology of molecular markers and waxy corn, which is applied in the field of biological identification, can solve the problems that the research has not yet been carried out, and achieve the effect of good repeatability, simple method, and less sample consumption
Active Publication Date: 2016-04-13
SHANGHAI ACAD OF AGRI SCI +1
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However, research on the variety identification and molecular basis of genetic diversity of Huwucaihuanuo 1 has not yet been carried out.
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Embodiment 1
[0059] Example 1 Determining SNP Molecular Markers
[0060] The SNP site sequence was obtained by 2b-RAD degenerate genome sequencing of Huwucaihuanuo 1, including 9 SNP site sequences SNP1-SNP9 of Huwucaihuanuo 1, and each SNP molecular marker sequence contained 1 Hu The SNP site of Wucaihuanuo 1. The nucleotide sequences of the molecular markers SNP1-SNP9 are respectively shown in SEQ ID NO.1-9, and the specific SNP molecular marker information is shown in Table 3.
Embodiment 2
[0061] Example 2 Variety identification of corn Huwucaihuanuo 1
[0062] Using the combination of primers designed in this invention to carry out variety identification on genomic DNA samples of glutinous rice varieties Huwucaihuanuo 1, Shen W48, Shen W93, Huziheinuo 1, Hucainuo 1, and Huyunuo 3 and identification of germplasm resources.
[0063] (1) Design primers
[0064] According to the SNP molecular markers shown in Table 3 determined in Example 1, a pair of AC-PCR primers is designed, and the AC-PCR primers include primer pair combinations 1-9, and each primer pair combination includes two primer pairs, specifically See Table 2 for primer sequences;
[0065] (2) Genomic DNA extraction
[0066] The CTAB method is used to extract the genomic DNA of any organ or tissue of the samples to be identified: sample 1, sample 2, sample 3, sample 4, sample 5, sample 6.
[0067] (3) PCR amplification
[0068] Perform PCR amplification on the genomic DNA of the sample to be ident...
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Abstract
Disclosed are an SNP molecular marker for identifying Shanghai colorful waxy corn NO.1 and an identifying method thereof. The identifying method comprises the steps of using a sequence of nine SNP sites obtained through degeneracy genomic sequencing of Shanghai colorful waxy corn NO.1 2b-RAD to design nine pairs of AC-PCR primers based on the sequence of the SNP sites; analyzing an obtained fingerprint spectrum through a PCR amplification technology by agarose gel electrophoresis; identifying whether the product is the Shanghai colorful waxy corn NO.1 according to patterns of electrophoretic bands of the nine SNP sites. The number of samples to be detected is small, the method is simple and quick, identified results are accurate, and good application and popularization prospects are achieved.
Description
technical field [0001] The invention belongs to the technical field of biological identification, and in particular relates to a SNP molecular marker for identifying waxy corn Huwucaihuanuo 1 and an identification method thereof. Background technique [0002] Waxy maize (Zeamays L. certainakulesh) is a special maize type with rich nutritional value formed by recessive mutation of maize endosperm wx gene. my country is the origin of waxy maize, and the local germplasm resources are abundant. According to statistics, more than 900 germplasms of waxy maize have been collected in the National Gene Bank of China. The identification of waxy maize varieties is not only helpful to the efficient utilization of maize germplasm resources, but also of great significance to the study of genetic diversity of maize lines. [0003] Single nucleotide polymorphism (singlenucleotide polymorphism, SNP) mainly refers to the DNA sequence polymorphism caused by the variation of a single nucleotid...
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