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Cotton blastogenesis identification method based on SSR markers and capillary electrophoresis

A technology of capillary electrophoresis and genetic identification, applied in the application of molecular marker-assisted selection breeding, based on SSR markers and capillary electrophoresis in the field of high-throughput genetic identification, can solve problems such as difficult analysis methods, and achieve high resolution and high degree of automation , The analysis results are accurate and reliable

Inactive Publication Date: 2016-07-06
XINJIANG ACADEMY OF AGRI & RECLAMATION SCI
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Problems solved by technology

[0006] The purpose of the present invention is to solve the problem of difficult cotton genetic identification and analysis methods under the current big data, and provide a method for genetic identification and analysis of cotton germplasm resources that can achieve high-throughput, large-scale, multi-batch, and high-efficiency. The results are accurate, reliable, economical and practical based on SSR markers and capillary electrophoresis cotton germplasm genetic identification method,

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  • Cotton blastogenesis identification method based on SSR markers and capillary electrophoresis
  • Cotton blastogenesis identification method based on SSR markers and capillary electrophoresis
  • Cotton blastogenesis identification method based on SSR markers and capillary electrophoresis

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Embodiment Construction

[0037] Refer to attached figure 1 , figure 2 , image 3 , 95 kinds of cotton varieties were selected as experimental materials. A method for genetic identification of cotton germplasm based on SSR markers and capillary electrophoresis, characterized in that it comprises the following steps:

[0038] 1. Cotton genomic DNA extraction, for the experimental material of above-mentioned 95 cotton varieties data, carry out the extraction of DNA respectively:

[0039] (1) Take 1 young cotton leaf and put it into a 1.5ml centrifuge tube, add 0.6ml extraction buffer to make 500mL formula as follows: 0.35MGlucose: 34.68g, 0.1MTris.HCl: 50mL, 5mMNa.EDTA: 5mL, 2 %PVP: 100mL, 1%β-Me: 5mL, ddWater: 340mL, grind with electric drill;

[0040] (2) Centrifuge at 10,000rpm (Eppendorf) for 15min (4°C), discard the supernatant; add 0.6ml of lysis buffer (500mL) preheated at 65°C to the pellet as follows: 1.4MNaCl: 40.908g, 0.1MTris.HCl: 50mL, 20mMNa.EDTA: 20mL, 2%CTAB: 100mL, 2%PVP: 100mL, 1%...

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Abstract

The invention belongs to thebiotechnology field, and relates to a genetic identification method based on SSR markers and capillary electrophoresis high throughput for cotton. The cotton blastogenesis identification method based on the SSR markers and capillary electrophoresis is characterized by comprising the following steps that 1, a plurality of tender cotton leaves are adopted as research materials, and cotton genomic DNA is extracted; 2, primers are screened and synthesized; 3, PCR amplified reaction is carried out; 4, capillary gel electrophoresis detection is carried out; 5, data statistics and analysis of genetic diversity is carried out. By means of the method, time and labor are saved, and electrophoresis film analysis automation and data processing computerization can be achieved, so that genetic identification analysis is carried out on cotton genetic resource materials with high throughout, large scale, multiple batches and high efficiency, and the result is accurate, reliable, economical and practical.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a high-throughput genetic identification method for cotton based on SSR markers and capillary electrophoresis. The method is suitable for genetic relationship identification of germplasm resources, variety purity and authenticity identification, and molecular marker assistance. Application for selection breeding. Background technique [0002] Cotton belongs to Malvaceae and Gossypium, and has 51 species in total, consisting of 4 cultivated species and 47 wild species. China is one of the countries with the largest cotton planting area. Among the five major cotton producing countries including China, the United States, India, Pakistan and Uzbekistan, China's cotton field area is in the top two or three. At present, there are 6,100 upland cotton germplasms, 387 sea-island cottons, 373 Asian cottons, 8 African cottons, 241 semi-wild germplasms of upland cotton, 31 wild species, etc., tot...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/6827C12Q1/6895C12Q2525/151C12Q2565/125
Inventor 董承光王娟李保成周小凤马晓梅余渝李生秀陈红王新王长彪李成奇
Owner XINJIANG ACADEMY OF AGRI & RECLAMATION SCI
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