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Rhododendron fortunei SSR primer pair based on transcriptome sequencing development, screening method and application

A technology of transcriptome sequencing and primer pairs, which is applied in biochemical equipment and methods, recombinant DNA technology, microbial measurement/inspection, etc., can solve the problems of limited application, and achieve the effect of fast method, high versatility and practicability

Active Publication Date: 2019-03-15
HUANGGANG NORMAL UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are few reports on the polymorphic SSR markers of Rhododendron versicolor, which greatly limits its application in genetic research.

Method used

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  • Rhododendron fortunei SSR primer pair based on transcriptome sequencing development, screening method and application
  • Rhododendron fortunei SSR primer pair based on transcriptome sequencing development, screening method and application
  • Rhododendron fortunei SSR primer pair based on transcriptome sequencing development, screening method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] [Example 1] Development of EST-SSR primers for Rhododendron versicolor with high polymorphism

[0039] Based on the high-throughput sequencing of transcriptome provided by the present invention, combined with bioinformatics methods for SSR sequence search and SSR marker primer design and verification, the specific implementation methods are as follows:

[0040] (1) Screening of Unigene sequences rich in SSR sites: Total RNA was extracted from Rhododendron brocade materials, a transcriptome sequencing library was constructed, and high-throughput sequencing was performed using Illumina next-generation sequencer. The sequencing data was assembled after strict filtering to obtain Unigene, which was used as background data for subsequent SSR primer development. Use MISA software to search for SSR sites on Unigene, the search criteria are: dinucleotides, trinucleotides, tetranucleotides, pentanucleotides, and hexanucleotides, the minimum repeat times are 10, 7, 6, respectivel...

Embodiment 2

[0046] [Example 2] Application of 25 pairs of EST-SSR primers in the study of genetic diversity of Rhododendron yunnanensis population

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Abstract

The invention discloses a rhododendron fortunei SSR primer pair based on transcriptome sequencing development, a screening method and application, and belongs to the technical field of biology. The primers are obtained on the basis of transcriptome sequencing and sequence analysis, on the basis of transcriptome sequencing, a large number of data are screened, a unigene sequence which is rich in SSR sites is acquired and SSR labeled primers are designed, and the barriers of a small number of rhododendron fortunei SSR molecular markers and low development efficiency at present are overcome. By arhododendron fortunei group, the effectiveness of the SSR primers is verified, and a foundation is laid for genetic research such as rhododendron fortunei genetic diversity research, genetic linkagemap, QTL positioning and molecular marker-assisted breeding.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a pair of SSR primers for Rhododendron yunnanensis developed based on transcriptome sequencing, a screening method and an application. Background technique [0002] Yunjin Rhododendron (Rhododendron fortunei Lindl.) is an evergreen shrub or small tree that mainly grows at an altitude of 600-2000m. It is a unique Rhododendron species in China. Rhododendron fortunei Lindl. The important ancestral parent of the hybrid, the important maintainer of the ecological function of the mountain top, is distributed in Shaanxi, Hunan, Zhejiang, Hubei, Jiangxi, Guangxi, Guizhou, Yunnan, Fujian and other provinces in my country. The flowers are fragrant and have strong resistance. High ornamental value and economic value. The research reports on Rhododendron versicolor mainly focus on the morphological characteristics of regional Rhododendron versicolor plants, resource distribution, reso...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12Q1/6869C12N15/11
CPCC12Q1/6869C12Q1/6895C12Q2600/156
Inventor 王书珍金卫斌张明菊李志良夏奥雷张羽佳黄诗颖
Owner HUANGGANG NORMAL UNIV
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