Preparation method and application for Miscanthus Genic-SSR mark

Abstract

The invention discloses a preparation method and application for a Miscanthus Genic-SSR mark. The method comprises the following steps: A, acquiring Miscanthus transcriptomic sequence, that is, selecting young and tender Miscanthus leaves, extracting total RNA respectively so as to obtain transcriptome Reads, removing the pollution of carrier RNA sequence and carrying out splicing so as to obtainthe Miscanthus transcriptomic sequence; B, identifying SSR sites in Miscanthus ESR sequence, that is, searching SSR sites in obtained non-redundant Unigene-EST sequence by using SSR retrieval software, with repeats being 2 to 6 bases, so as to obtain a series of Unigene sequences containing information about the SSR sites; C, designing Miscanthus Genic-SSR primers, that is, using primer design software to design primers according to the obtained Unigene sequences containing the SSR sites so as to obtain a series of the Miscanthus Genic-SSR primers. According to the invention, the method is simple, has high efficiency and is simple to operate; a huge amount of marks are obtained; EST sequence obtained through transcriptomic sequencing lays a foundation for genetic research, molecular improvement and the like of the critical energy plant Chinese silvergrass.

Description

technical field [0001] The invention belongs to the technical field of molecular biology, and more specifically relates to a preparation method of a Miscanthus Genic-SSR marker, and also relates to an application of a Miscanth Genic-SSR marker. technical background [0002] Miscanthus belongs to the C4 perennial herbaceous plants of Poaceae, mainly including Miscanthus, Wujiemans, Di, Nandi, and Qigang, which are concentrated in the east and southeast of Asia. Miscanth plants have the characteristics of strong vitality, high photosynthetic utilization efficiency, and strong stress resistance, and can be used in pharmaceuticals, papermaking, water and soil conservation, and ecological restoration. With the continuous improvement of human energy demand and the continuous reduction of global energy reserves, Miscanthus has attracted more and more attention as a potential energy plant. At present, Qigang, which has been widely planted in Europe and the United States, has an ann...

AI Technical Summary

Problems solved by technology

Among them, the first two methods are time-consuming, laborious and costly to develop, and the third method relies on the existing EST data in NCBI, and its application to species with few EST sequencing is very limited.

[0004] As of November 21, 2011, there are only 5 EST sequences of Miscanthus that can be retrieved in the NCBI database, which is difficult to meet the needs of developing EST-SSR markers; and in the existing research, the development of molecular markers of Miscanthus only Limited to the use of magnetic beads or the establishment of library mining, it cannot meet the current research needs of Miscanthus. Therefore, obtaining a large amount of EST sequence information through transcriptome sequencing and using bioinformatics to mine SSR markers has become an efficient technical means

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