Microsatellite marker site development method and microsatellite marker length detection method within a microsatellite marker site

Abstract

The invention discloses a method for developing microsatellite marker sites, a method for detecting the length of microsatellite markers in the microsatellite marker sites, and a probe set for development. The development method includes: obtaining a mixed sample; extracting the genome of the mixed sample; fragmenting the genome to obtain genome fragments; using the probe set to hybridize with the genome fragments respectively; purifying the genome fragments successfully hybridized in multiple hybridization solutions; After mixing the purified hybrid genome fragments, use high-throughput sequencing to detect the purified genome fragments; obtain effective high-throughput sequencing fragments; and classify the effective high-throughput sequencing fragments. The detection method comprises: selecting the microsatellite marker site to be detected; using multiple amplification primers to amplify the microsatellite markers in the microsatellite marker site to be detected to obtain the microsatellite marker site in the microsatellite marker site The length of the satellite marker. The above method is simple, fast, comprehensive and accurate.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for developing microsatellite marker sites and a method for detecting the length of microsatellite markers in the microsatellite marker sites. Background technique [0002] Microsatellite markers are also called short tandem repeats (short tandem repeats, STR) or simple sequence repeats (simple sequence repeats, SSR), which refer to tandem repeats consisting of more than two nucleotides as repeating units. Microsatellite marker loci refer to loci containing microsatellite markers on the genome. Microsatellite marker loci are abundant and evenly distributed on the genome. The development of microsatellite marker loci refers to the search for microsatellite marker loci on the genome point process. In different samples, the repeating times of microsatellite marker repeat units in the same microsatellite marker locus may be different, and there are length variations between sam...

AI Technical Summary

Problems solved by technology

[0005] The development and detection process of microsatellite marker sites is complicated, the throughput is low, and it is extremely time-consuming and laborious; secondly, the resolution of electrophoretic detection of microsatellite marker sites is low, and the detection results are inaccurate. Accurate results need to be corrected by reference samples, etc.

The problems derived from this include: there are few microsatellite marker sites developed, usually less than 200, accounting for about 1% of all microsatellite marker sites on the genome; the samples used to test the polymorphism of microsatellite marker sites are also limited. There are few, usually around tens, so the polymorphism detection results are inaccurate; the flanking sequence conservation of microsatellite marker sites is unknown, which affects the versatility of primers for amplifying microsatellite marker sites; the detection of microsatellite markers The number of loci is limited. Generally, dozens of microsatellite marker loci are detected in a sample to be tested, resulting in incomplete and inaccurate DNA ID card information of the established sample.

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