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The ssr primer pair, screening method and application of rhododendron yunnanensis developed based on transcriptome sequencing

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

Active Publication Date: 2021-09-24
HUANGGANG NORMAL UNIV
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  • Summary
  • 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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  • The ssr primer pair, screening method and application of rhododendron yunnanensis developed based on transcriptome sequencing
  • The ssr primer pair, screening method and application of rhododendron yunnanensis developed based on transcriptome sequencing
  • The ssr primer pair, screening method and application of rhododendron yunnanensis developed based on transcriptome sequencing

Examples

Experimental program
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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. MISA software was used to search for SSR sites in Unigene. The search criteria were: dinucleotides, trinucleotides, tetranucleotides, pentanucleotides, and hexanucleotides, the minimum repeat times were 10, 7, 6, 5, ...

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 pair of SSR primers for Rhododendron versicolor developed based on transcriptome sequencing and a screening method and application. Belongs to the field of biotechnology. The primers are obtained based on transcriptome sequencing and sequence analysis. On the basis of transcriptome sequencing, a large amount of data is screened to obtain unigene sequences rich in SSR sites and design primers for SSR markers, which overcomes the current SSR molecular The obstacles of small number of markers and low development efficiency. The validity of the SSR primers was verified by Rhododendron versicolor population, which laid a foundation for the genetic diversity research, genetic linkage map structure, QTL mapping, molecular marker-assisted breeding and other genetic research of Rhododendron versicolor.

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 Patents(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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