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Method for developing genome simple sequence repeats (SSR) molecular marker

A molecular marker and genome technology, applied in the field of molecular biology, can solve the problems of difficult operation, primer-specific reading errors, time-consuming, etc., and achieve the effect of improving efficiency, reducing human and financial resources, and reducing sequencing costs.

Inactive Publication Date: 2015-01-28
JIANGSU UNIV
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Problems solved by technology

Pre-screening generally uses polyacrylamide gel electrophoresis or capillary electrophoresis, but polyacrylamide gel electrophoresis is a relatively difficult experimental technique, which is time-consuming, and it is easy to produce artificial readings when the specificity of the primers is not good. There are errors; in addition, although capillary electrophoresis is convenient and highly accurate, it is also expensive

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  • Method for developing genome simple sequence repeats (SSR) molecular marker
  • Method for developing genome simple sequence repeats (SSR) molecular marker
  • Method for developing genome simple sequence repeats (SSR) molecular marker

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Embodiment Construction

[0031] In order to make the object, technical solution and advantages of the present invention clearer, the present invention will be further described in detail below in conjunction with the accompanying drawings and embodiments. It should be understood that the specific embodiments described here are only used to explain the present invention, not to limit the present invention.

[0032] Materials: Colocasia gigantea was selected as the test sample, which has no genome and transcriptome except a few chloroplast genes in GeneBank. Take two wild taro plants, one from Xishuangbanna, Yunnan Province, China, and the other from Vietnam. See flow chart figure 1 .

[0033] method:

[0034] (1) DNA extraction: cut 0.4 g of fresh and tender leaves and extract DNA with the CTAB method, and then use agarose gel electrophoresis to detect the extraction effect, such as figure 2 As shown in , samples with clear main band and no smear can be used for subsequent library construction ope...

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Abstract

The invention discloses a method for developing genome simple sequence repeats (SSR) molecular marker. The method comprises the steps of taking two plant samples, respectively establishing banks for DNA of the samples, thereby obtaining two corresponding libraries; respectively sequencing the libraries by an Illumina sequencing technology, then carrying out micropackaging, seeking in Contigs obtained through micropackage so as to obtain the SSR of the samples, and screening the common SSR in the two samples, wherein the common SSR has the same chromosomal location and the same repetitive unit; and carrying out polymorphism screening on the two samples according to the common SSR, and judging whether the two samples are in porlymorphism according to the difference of times of the repetition of the SSR of the two samples. The method not only meets the requirement of seeking a lot of SSR molecular markers, but also has low cost; due to the advantages of high polymorphism, dominant heredity, wide distribution and few template DNA, the SSR molecular markers can be widely used in genetic diversity analysis, genetic mapping, quantitative trait loci (QTL) location, molecular marker assisted breeding and the like.

Description

technical field [0001] The invention belongs to the field of molecular biology, and in particular relates to a technology related to the development of SSR molecular markers in a genome sequencing library. Background technique [0002] DNA molecular markers refer to specific DNA fragments that can reflect certain differences in the genomes of organisms or populations. Compared with conventional morphological markers, cytological markers and biochemical markers, DNA molecular markers directly have the following advantages: (1) Exist in any growth stage of animals and plants in the form of DNA, and can be detected in any tissue , not limited by seasons and environments, and there is no problem of expression; (2) the number is extremely large, the genome variation is extremely rich, the number of markers is almost unlimited, the polymorphism is high, and a large number of primers and probes can be used to complete Analysis covering the genome; (3) Many markers are co-dominant,...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/6869
Inventor 刘宇婧郭钰龙春林吴沿友付为国邢德科张川
Owner JIANGSU UNIV
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