The Manshan Red SSR primer based on the transcription group sequencing method and screening method and application
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A technology of transcriptome sequencing and Manshanhong, which is applied in biochemical equipment and methods, recombinant DNA technology, microbial measurement/testing, etc., to achieve the effect of fast method, high versatility and practicability
Active Publication Date: 2021-09-24
HUANGGANG NORMAL UNIV
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[0004] With the continuous development and maturity of the second-generation high-throughput sequencing technology, it has become possible to obtain large-scale EST-SSR molecular markers through transcriptome sequencing, but there are no reports on the development of SSR using the transcriptome data of Manshanhong.
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Embodiment 1
[0037] [Example 1] Development of EST-SSR primers of Manshanhong with high polymorphism
[0038] 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:
[0039] (1) Screening of Unigene sequences rich in SSR sites: The petal materials in the full-blooming stage of Mangosaurus were selected to extract total RNA, and a transcriptome sequencing library was constructed, and high-throughput sequencing was performed using Illumina next-generation sequencer. Strictly filter the sequencing data and assemble the transcriptome data to obtain Unigene, which is used as the background data for the development of SSR markers. Use MISA software to search for SSR sites on Unigene sequences. The search criteria are: dinucleotides, trinucleotides, tetranucleotides, pentanucleotides, and hexanucleotide...
Embodiment 2
[0045] [Example 2] Application of 25 pairs of EST-SSR primers in the study of genetic diversity of Manshanhong population
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Abstract
The invention discloses a pair of SSR primers 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 EST‑SSR markers, which overcomes the current problems The low number and low development efficiency of red SSR molecular markers are obstacles. The validity of the EST-SSR primers was verified by the Mangospermum population, which laid a foundation for the genetic diversity research, genetic linkage map structure, QTL mapping, molecular marker-assisted breeding and other genetic studies of Mangosteen.
Description
technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a pair of SSR primers developed based on transcriptome sequencing and a screening method and application. Background technique [0002] Manshanhong (Rhododendron mariesii Hemsl.et Wils) is a deciduous shrub of the family Ericaceae (Rhododendron). It is mainly distributed in Hebei, Shaanxi, Hubei, Jiangsu, Anhui, Jiangxi, Fujian, Taiwan, Henan, Zhejiang, Hunan, Guangdong, Guangxi, Sichuan, Guizhou and other places have high horticultural ornamental value. Manshanhong has the effects of relieving cough, eliminating phlegm and relieving asthma, and is used to treat acute and chronic bronchitis, asthma and other diseases. The chemical components that have been isolated from Manshanhong mainly include flavonoids and volatile oils, such as hyperin, rhododendron, kaempferol, quercetin, myricetin, and Huangmeisu. For a long time, many breeding scholars have carried out in-depth...
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