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Genetic analysis method based on capillary electrophoresis and SSR fluorescence labeling for rice

A genetic analysis method and capillary electrophoresis technology, applied in biochemical equipment and methods, material analysis by electromagnetic means, analysis of materials, etc., can solve difficult to achieve, difficult to integrate and accurately compare DNA data, and unable to accurately read fragments. size and other issues, to achieve the effect of reducing human error, accurate experimental results, and reducing workload

Inactive Publication Date: 2014-07-09
广西益谱检测技术有限公司 +3
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Problems solved by technology

At present, the most commonly used method is polyacrylamide gel electrophoresis detection method. This method can theoretically separate two fragments with a difference of 1bp, but it is difficult to achieve in practice, and the fragment size cannot be read accurately. It can only be estimated artificially, which is difficult Effective integration and accurate comparison of large-scale DNA data of different batches

Method used

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  • Genetic analysis method based on capillary electrophoresis and SSR fluorescence labeling for rice
  • Genetic analysis method based on capillary electrophoresis and SSR fluorescence labeling for rice
  • Genetic analysis method based on capillary electrophoresis and SSR fluorescence labeling for rice

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0029] Example: UPGMA Cluster Analysis of Rice Varieties

[0030] 18 commonly used rice varieties were selected as experimental materials

[0031] 1. Rice Genomic DNA Extraction

[0032] (1) Put the rice seeds at a constant temperature of 30°C to germinate and emerge. When 2-3 true leaves grow, take the young rice leaves in a mortar, add liquid nitrogen to grind them into powder, and put them into a 2 ml centrifuge tube ;

[0033] (2) Add 1.2 ml 65°C preheated SDS extraction solution (Tris-HCl 200 mmol / L, pH8.0; EDTA 50 mmol / L pH8.0; NaCl 500 mmol / L; 3% SDS) into the centrifuge tube , Shake the sample powder and the extract quickly to prevent them from forming agglomerates, put in a water bath at 65°C for 45 minutes, and shake gently 4-6 times during the period;

[0034](3) Add 200 μl KAc (5 mol / L, pH 4.8), mix well, and place in ice bath for 15 min;

[0035] (4) Centrifuge at 10,000 rpm for 15 min in a refrigerated centrifuge at 4°C;

[0036] (5) Take 800 μl of supernata...

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Abstract

The invention provides a genetic analysis method used for rice. According to the method, capillary gel electrophoresis is utilized to detect SSR-PCR amplification products, and the high resolution characteristic of the capillary gel electrophoresis technology is utilized to distinguish rice varieties with near genetic relationship, so that analysis results are more accurate. By adoption of the genetic analysis method based on capillary gel electrophoresis and SSR labeling, a dendrogram established by UPGMA clustering analysis is used for genetic relationship analysis of rice varieties. The genetic analysis method has advantages of high resolution, rapidness, high efficiency, accurate and reliable results, and the like. The method has characteristics of easy automation and convenience for integration and analysis of a large amount of data, which are beyond the grasp of traditional detection methods.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a genetic analysis method for rice based on capillary electrophoresis technology and SSR fluorescent markers. The method is also applicable to collection and management of germplasm, identification between varieties, determination of parent-child relationship and Development of molecular marker-assisted selection breeding programs. Background technique [0002] The genus Oryza, belonging to the rice family (Oryzeae Dumort.), consists of 2 cultivated species and 20 wild species, widely distributed in tropical and subtropical regions. The genus Oryza is very rich in genetic resources. It is estimated that there are 140,000 unique germplasms of rice cultivated in Asia alone in the world. In China, with the large-scale promotion of hybrid rice and the successful application of super rice, the yield of rice has been greatly improved. Therefore, the study of genetic diversity of rice and th...

Claims

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Application Information

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IPC IPC(8): C12Q1/68G01N27/447
CPCC12Q1/6858C12Q2531/113C12Q2565/125C12Q2563/107
Inventor 梁俊陈远孟刘守廷覃红萍甘国勇莫璋红宋雪萍蒋天成赵晶苏小玲刘广林陈传华唐梅张宗琼罗群昌梁益珍李碧红
Owner 广西益谱检测技术有限公司
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