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Preparation method and use of simple sequence repeat (SSR) marker for screening miscanthus by magnetic bead enrichment process

A technology of magnetic beads and awn

Inactive Publication Date: 2011-06-01
湖北光芒能源植物有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there are few studies on the genetics of the five-jointed awn, especially at the DNA molecular level.

Method used

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  • Preparation method and use of simple sequence repeat (SSR) marker for screening miscanthus by magnetic bead enrichment process
  • Preparation method and use of simple sequence repeat (SSR) marker for screening miscanthus by magnetic bead enrichment process
  • Preparation method and use of simple sequence repeat (SSR) marker for screening miscanthus by magnetic bead enrichment process

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Embodiment 1

[0069] The present invention is further described below in conjunction with description:

[0070] A preparation method for screening Miscanthus plant SSR markers by magnetic bead enrichment method, the preparation steps are:

[0071] 1. Test materials: 66 plants of 5 species of Miscanthus as shown in Table 1, including 47 Miscanthus, 7 Miscanthus, 7 Grass, 3 Nandi, and 2 Qigang.

[0072] 2. Total DNA extraction method: extract the total DNA from the young leaves of the above materials. The specific method refers to Gu Hongya et al. (Gu Hongya, Qu Lijia. Experimental Handbook of Plant Molecular Biology [M]. Beijing: Gao et al. Educational Press, 19983~12.) Extraction method reported.

[0073]3. Hybridization of the probe with the target fragment, the specific method is as follows:

[0074] (1) Acquisition of Genomic DNA Fragment of Miscanthus pentaphyllum: Take about 125ng of total genomic DNA (5μL) and digest it in a 20μL enzyme digestion system (DNA: 5μL; 10x Buffer R: 5μL;...

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Abstract

The invention discloses a preparation method and use of a simple sequence repeat (SSR) marker for screening miscanthus by a magnetic bead enrichment process, which comprises: 1) extracting genomic DNA of miscanthus floridulus which belongs to miscanthus; 2) incising the extracted total DNA by using restriction incision enzyme to obtain a genomic DNA; 3) intercrossing a probe and a target fragment; 4) enriching the hybrid DNA molecules having an SSR sequence by using magnetic beads; 5) amplifying and purifying the DNA fragment having the SSR sequence; 6) cloning and sequencing the DNA fragment; and 7) designing an SSR primer according to the flanking sequence of SSR, and designing a specific primer by using primer design software for amplifying the satellite fragment at the locus. The method is easy and convenient for operation; meanwhile, the SSR marker has the characteristics of high codominance, high polymorphism, multiple allele property and the like and can be used for analyzing the genetic diversity and genetic relationship of miscanthus and used in construction of genetic map of miscanthus, calibration of a target gene and drawing of a fingerprint.

Description

technical field [0001] The invention belongs to the technical field of Miscanthus plant breeding, and more specifically relates to a preparation method for screening Miscanthus plant SSR markers by a magnetic bead enrichment method, and also relates to an application of the Miscanthus plant SSR markers screened by the magnetic bead enrichment method. Background technique [0002] Miscanthus, commonly known as Miscanthus, belongs to the tall perennial grasses of Poaceae. Since Miscanthus is not a food crop and it mainly grows on wasteland, Miscanthus as an energy plant has received widespread attention worldwide. Various countries have introduced corresponding policies and measures in order to be at the forefront in the development and research of energy plants. When U.S. President Barack Obama unveiled a new strategy aimed at reviving U.S. biofuel production, he "vowed never to let China and other countries take the lead in building a new energy economy." China is the dist...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 刁英胡小虎胡中立郑兴飞游永宁余作平
Owner 湖北光芒能源植物有限公司
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