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A kind of screening method and application of ssr molecular marker of Acanthopanax target gene

A screening method and molecular marker technology, applied in biochemical equipment and methods, microbe determination/inspection, DNA/RNA fragments, etc., to speed up the breeding process and overcome uncertainty

Active Publication Date: 2022-03-04
DALIAN NATIONALITIES UNIVERSITY
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Problems solved by technology

At present, there is no report on the development of SSR molecular markers for the target gene of Acanthopanax based on RNA-seq technology

Method used

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  • A kind of screening method and application of ssr molecular marker of Acanthopanax target gene
  • A kind of screening method and application of ssr molecular marker of Acanthopanax target gene
  • A kind of screening method and application of ssr molecular marker of Acanthopanax target gene

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Embodiment

[0017] 1. Extract total RNA from Acanthopanax fruit, root, stem and leaf

[0018] One Acanthopanax germplasm SL1 from Suileng County, Heilongjiang Province was used as material. The total RNA of Acanthopanax senticosus from different tissues and organs was extracted according to the method recommended by Shanghai Shenggong Plant RNA Extraction Kit. The total RNA concentration of each tissue and organ was 200-400ng / μL, and stored at -80°C.

[0019] 2. RNA-seq data analysis of Acanthopanax senticosus fruit, root, stem and leaf

[0020] A transcriptome library was constructed by reverse transcription using samples mixed with equal amounts of total RNA from four tissues and organs as templates. Clean reads were obtained by removing adapters, N ratio greater than 0.1% and low-quality reads from the raw reads obtained by sequencing, and the clean reads were spliced ​​using Trinity software to obtain unigene. Compare unigenes with known functional genes in public databases (Nr, Sw...

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Abstract

The invention discloses a screening method and application of the SSR molecular marker of the target gene of Acanthopanax. The specific steps of the screening method are: 1) extracting the total RNA of each tissue of Acanthopanax anticana; 2) performing RNA- Seq sequencing, Trinity software for unigene assembly; 3) Screen out unigenes related to the synthesis and accumulation of eleutherosides, SOD, etc., and MicroSatellite software screens the SSR sites on these unigenes; 4) Primer3.0 software designs primers and synthesizes them; 5) Genomic DNA is used as a template for PCR amplification; 6) PCR products are detected by electrophoresis, and SSR primers for the target gene are developed. The invention efficiently and more targetedly develops candidate SSR molecular markers related to target traits, and provides effective markers for molecular identification of excellent Acanthopanax germplasm resources. It is of great significance to the early identification of excellent Acanthopanax germplasm resources and to speed up the breeding process.

Description

technical field [0001] The invention belongs to the field of Acanthopanax senticosus molecular marker development and molecular biology technology, and relates to a screening method and application of Acanthopanax senticosus target gene SSR molecular marker. Background technique [0002] Acanthopanax is a deciduous shrub belonging to Araliaceae. The fruit is rich in superoxide dismutase (SOD), which has good free radical scavenging and antioxidant functions; the root bark is rich in saponins, tannins, linolenic acid and various vitamins etc. Both the fruit and the root can be used as medicine, which has the effects of enhancing human immunity, dispelling rheumatism, and strengthening bones and muscles. Acanthopanax is mainly distributed in Heilongjiang, Jilin, Liaoning, Hebei and Shanxi. In recent years, its planting area has gradually expanded. In the cultivation and promotion of Acanthopanax senticosus, the fruit quality of Acanthopanax senticosus is uneven, and the mole...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895C12Q2600/13C12Q2600/156
Inventor 丁健阮成江李雪柔韩平杨伞伞
Owner DALIAN NATIONALITIES UNIVERSITY
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