Exploitation method of orchid microsatellite marker site, and detection method of length of microsatellite marker in microsatellite marker site

Abstract

The invention discloses an exploitation method of an orchid microsatellite marker site, and a detection method of the length of a microsatellite marker in the microsatellite marker site. The exploitation method comprises the following steps: obtaining a mixed sample; extracting the genome of the mixed sample; fragmenting the genome to obtain genome fragments; respectively hybridizing a probe group with the genome fragments; purifying the successfully hybridized genome fragments in a hybridization solution; mixing the purified hybridized genome fragments, and detecting the purified genome fragments through using high flux sequencing; obtaining effective high flux sequencing fragments; and classifying the effective high flux sequencing fragments. The detection method comprises the following steps: selecting a microsatellite marker site to be detected; and amplifying the microsatellite marker in the microsatellite marker site to be detected by using a multiple amplification primer to obtain the length of the microsatellite marker in the microsatellite marker site. The methods are simple, fast, comprehensive and accurate.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for developing orchid 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 betw...

AI Technical Summary

Problems solved by technology

[0005] The development and detection process of microsatellite marker sites in orchids is complex, low-throughput, and 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 require reference samples, etc. Correction

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

Images