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Rice fingerprint constructing method based on SSR marking and capillary electrophoresis technology

A technology of capillary electrophoresis and fingerprinting, which is applied in the field of biological information, can solve the problems of inability to accurately read the size of amplified fragments, difficulty in uniformly processing reaction data, non-automation, etc., and achieve the effect of easy operation, avoiding human errors, and simple operation

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

[0004] Traditional SSR-PCR amplification products are mainly analyzed for polymorphism by biotechnology such as agarose gel electrophoresis or polypropylene gel electrophoresis. Although these methods are relatively mature, they are not automated, time-consuming, labor-intensive, and cannot Accurately read the size of the amplified fragments, it is difficult to uniformly process the reaction data of different batches, and it is still quite difficult to collect and analyze data in large-scale and multiple batches

Method used

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  • Rice fingerprint constructing method based on SSR marking and capillary electrophoresis technology
  • Rice fingerprint constructing method based on SSR marking and capillary electrophoresis technology
  • Rice fingerprint constructing method based on SSR marking and capillary electrophoresis technology

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Experimental program
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Embodiment

[0026] Example: Construction of rice variety SSR fingerprints

[0027] The experimental materials were selected 22 commonly used rice varieties.

[0028] 1. Genomic DNA extraction

[0029] (1) Scissors scrubbed with 75% alcohol and cut 0.02~0.04 cm 2Put the leaves of the leaves into the wells of a 96-well PCR plate, add 100 μl of DNA extraction buffer, add 0.2 μl of 100 mg / ml proteinase K and 1 μl of 10 mg / ml ribonuclease, and cover with a PCR plate silica gel cover Mix on a vortex for 10 sec;

[0030] (2) Set and save the following reaction program in the PCR instrument: first set the temperature to 37°C and the time to 30 min; then set the temperature to 68°C and the time to 20 min; then set the temperature to 37°C and the time to 30 min min; finally set the temperature to 4°C and the time to infinity;

[0031] (3) Put the PCR plate obtained in step (1) into the set PCR machine for reaction;

[0032] (4) Immediately after the reaction, transfer the PCR plate to ice for ...

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Abstract

The invention provides a rice fingerprint constructing method. The method includes: extracting rice genome DNA, screening SSR fluorescent primers with high polymorphism, subjecting the rice variety DNA to PCR amplification, detecting the PCR product by utilization of capillary gel electrophoresis, analyzing data to obtain a 0-1 matrix graph and converting the 0-1 matrix graph into a fingerprint mode mapping. By adoption of the characteristics of the SSR polymorphism and the high resolution of the capillary gel electrophoresis technology, the method can recognize two polymorphism bands with a band difference being 1 bp. The method is prone to automation, simple in operation, stable and reliable in results, and prone to large-scale multi-batch data integration and analysis, and is suitable for construction of a large number of rice variety fingerprints.

Description

technical field [0001] The invention relates to the technical field of biological information, in particular to a method for constructing rice fingerprints based on SSR fluorescent labeling and capillary electrophoresis technology. Background technique [0002] Rice is one of the main food crops in my country, and it is also one of the first batch of protected crops covered by the Regulations on the Protection of New Varieties of Plants in my country. The authenticity of rice seedlings is an important index to measure the quality of seedlings, which directly affects the yield, quality and economic benefits of rice. At present, rice seeds and seedlings in my country have the phenomenon of low purity, mixed quality, and shoddy quality. Crop variety approval agencies at all levels and seed and seedling production departments are paying more and more attention to the identification of fake and inferior rice varieties. However, the morphological characteristics of rice seed resou...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6858
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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