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Litopenaeus vannamei osmotic regulation related functional gene EST-SSR markers and specific primers and detection method thereof

A technology of osmotic pressure regulation and related functions, applied in the field of molecular markers, which can solve the problems of less EST-SSR marker development and no EST-SSR marker development yet.

Active Publication Date: 2016-09-28
SOUTH CHINA SEA INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, so far, few EST-SSR markers have been developed in Litopenaeus vannamei, and no EST-SSR markers associated with specific functional genes have been developed.

Method used

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  • Litopenaeus vannamei osmotic regulation related functional gene EST-SSR markers and specific primers and detection method thereof

Examples

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Embodiment 1

[0030] 1. Transcriptome sequencing and sequence alignment of Litopenaeus vannamei

[0031] 1.1 Transcriptome sequencing

[0032] The muscles, hepatopancreas, gills and intestines of Litopenaeus vannamei were collected, and the total RNA was extracted with RNAiso Plus (TaKaRa, Japan) and treated with DNase I (TaKaRa). Using NanoDrop TM2000 Spectrophotometer (Thermo Scientific Waltham, MA, USA) was used to measure RNA concentration, and Agilent 2100 Bioanalyzer (Agilent Technologies, CA, USA) was used to evaluate the quality of RNA. After equal amounts of RNA extracted from four different tissue samples were mixed, a cDNA library was established through processes such as mRNA enrichment, mRNA fragmentation, cDNA synthesis, end repair, "A" tailing, and sequencing adapter ligation. The cDNA library was sequenced using the IIIlumina HiSeq2000 high-throughput sequencing platform, and seqClean and Lucy software were used to remove adapter sequences and low-quality sequences to obta...

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Abstract

The invention discloses litopenaeus vannamei osmotic regulation related functional gene EST-SSR markers and specific primers and a detection method thereof. Litopenaeus vannamei genome DNA serves as a template, the specific primers of the litopenaeus vannamei osmotic regulation related functional gene EST-SSR markers are used for conducting PCR amplification on microsatellite locus sequences, and then typing is conducted on PCR amplification products through a sequenator. The specific primers and detection method of the litopenaeus vannamei osmotic regulation related functional gene EST-SSR markers can be used for population genetic structure analysis and molecular marker assisted breeding of litopenaeus vannamei and particularly have an important application value in breeding of good salt-tolerant varieties of the litopenaeus vannamei.

Description

Technical field: [0001] The invention belongs to the technical field of molecular markers, and in particular relates to EST-SSR markers of functional genes related to osmotic pressure regulation of Litopenaeus vannamei, specific primers and detection methods thereof. Background technique: [0002] Litopenaeus vannamei, commonly known as Penaeus vannamei, is native to South America and is distributed from the southwestern coast of Mexico to the Pacific coast in western Peru. It is the largest farmed prawn in the world. In my country, Litopenaeus vannamei aquaculture areas are spread all over the country's coastal marine aquaculture areas and inland freshwater aquaculture areas, and it is a pillar industry of aquaculture in my country. Litopenaeus vannamei introduced and bred in my country are all artificially bred populations, which are prone to genetic decline. Therefore, the selection and breeding of improved species is a major issue related to the success or failure of Li...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6888C12Q2600/156C12N15/11C12Q1/68
Inventor 任春华江晓陈廷黄文胡超群
Owner SOUTH CHINA SEA INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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