Single nucleotide polymorphism based primer set for hybrid identification of true and false peanuts and identification method thereof

A single nucleotide polymorphism and primer set technology, applied in biochemical equipment and methods, microbial measurement/testing, recombinant DNA technology, etc., can solve inaccurate detection results, low resolution of electrophoresis detection, and accurate identification results Reduce the speed and other problems, to save time and improve efficiency

Active Publication Date: 2018-04-20
石家庄博瑞迪生物技术有限公司
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Problems solved by technology

Taking a hybrid combination as an example, 5%-10% polymorphism among peanut materials is calculated. This hybrid combination needs to screen 50-60 markers to obtain enough differential markers, and then use the screened markers to test the F1 hybrids. It takes two days to complete the whole experiment; ②The electrophoretic detection resolution of microsatellite markers is low, an

Method used

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  • Single nucleotide polymorphism based primer set for hybrid identification of true and false peanuts and identification method thereof
  • Single nucleotide polymorphism based primer set for hybrid identification of true and false peanuts and identification method thereof
  • Single nucleotide polymorphism based primer set for hybrid identification of true and false peanuts and identification method thereof

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Experimental program
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Embodiment Construction

[0042] Below in conjunction with embodiment the present invention is further elaborated.

[0043] 1. Selection of materials: 21 peanuts were selected, and the genomic DNA of the peanut samples was extracted using the BioTeke Magnetic Bead Method Plant Genomic DNA Extraction Kit (Cat#AU31111).

[0044] 2. Using peanut genomic DNA as a template, a round of PCR amplification was performed with the primer mixture to obtain the target region;

[0045] The one-round PCR amplification system: primer mixture 10 μl; DNA dosage 100 ng; 3*M enzyme 15 μl; add water to make up 45 μl.

[0046] The one-round PCR amplification system: 95°C for 3min; (95°C for 30s, 60°C for 4min, 72°C for 30s), 17 cycles; 72°C for 4min.

[0047]Primer mixture: there are a total of 136 primers in the present invention, as shown in Table 1. Take 10 μl of each primer and dilute to 10 ml. The concentration of each primer is 0.1 μM.

[0048] Table 1 Primer Set

[0049]

[0050]

[0051]

[0052]

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Abstract

The invention relates to a single nucleotide polymorphism based primer set for hybrid identification of true and false peanuts and a method for performing hybrid identification on the true and false peanuts through the same primer set. With the adoption of the primer set and the method, 68 bites of up to 384 samples can be detected within one day, and the efficiency is increased 50-100 times by being compared with that of a microsatellite detection method. According to the primer set and the method, the single nucleotide polymorphism is directly read through the sequencing technique; the result is clear and reliable, and free from inter-sample interference problem.

Description

technical field [0001] The invention relates to a single nucleotide polymorphism peanut true and false hybrid identification primer set and method thereof. Background technique [0002] Peanut (Arachis hypogaea L.) is the fourth largest oil crop in the world and an important oil crop in the tropics and subtropics. It has been planted in more than 100 countries, and China is the largest peanut producer in the world. Peanut is a strictly self-pollinating crop. In the process of hybrid breeding, it is necessary to manually complete the whole process of female parent pollen detasseling, male parent pollen collection, and pollination. There are many uncontrollable factors in the process, resulting in a high probability of hybrids producing false hybrids. . Timely and accurate identification of true and false hybrids in the hybrid F1 generation is the key to the success of hybrid breeding. The traditional identification of true and false hybrids is mainly carried out by two meth...

Claims

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Application Information

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IPC IPC(8): C12Q1/6895C12Q1/6869C12N15/11
CPCC12Q1/6869C12Q1/6895C12Q2600/156
Inventor 张嘉楠梁炫强陈小平鲁清李海芬陶家军许彦芬李凝张萌
Owner 石家庄博瑞迪生物技术有限公司
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