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Primer sequence for detecting purity of hybrid seeds of Jinyou No.35 cucumbers and detection method

A technology of hybrid seed purity and hybrid seed, applied in the direction of DNA / RNA fragments, recombinant DNA technology, etc., to achieve fast and accurate identification results, low cost, and labor-saving effects

Inactive Publication Date: 2011-08-31
TIANJIN RES INST OF VEGETABLE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The SSR identification method of cucumber seed purity has not been reported yet

Method used

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  • Primer sequence for detecting purity of hybrid seeds of Jinyou No.35 cucumbers and detection method
  • Primer sequence for detecting purity of hybrid seeds of Jinyou No.35 cucumbers and detection method

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Effect test

Embodiment 1

[0017] The method for detecting the purity of Jinyou No. 35 cucumber hybrid seeds comprises the steps:

[0018] (1) Extracting the genomic DNA of the root tip of Jinyou 35 cucumber hybrid seeds germinated for 35 hours;

[0019] (2) Carry out PCR amplification: Add Jinyou No. 35 cucumber hybrid seed genomic DNA 10ng in the special thin-walled tube for PCR amplification, then add 15ng of the nucleotide sequence described in SEQ ID NO.1 in the sequence listing, and the sequence listing 15ng of the nucleotide sequence described in SEQ ID NO.2, 1 times of Mg 2+ PCR buffer solution, dNTP 2mmol, Taq DNA polymerase 0.5 unit, add sterile double distilled water to 10μl; put the special thin-walled tube for PCR amplification into the PCR instrument for amplification, the amplification conditions are: 94°C pre-denaturation for 180 seconds ; Denaturation at 94°C for 30 seconds, annealing at 50°C for 30 seconds, extension at 72°C for 60 seconds, 30 cycles; extension at 72°C for 7 minutes, ...

Embodiment 2

[0023] The method for detecting the purity of Jinyou No. 35 cucumber hybrid seeds comprises the steps:

[0024] (1) Genomic DNA was extracted from cucumber hybrid seeds of Jinyou 35;

[0025] (2) Perform PCR amplification: add 20ng of Jinyou 35 cucumber hybrid seed genomic DNA in the thin-walled tube dedicated to PCR amplification, then add 30ng of the nucleotide sequence described in SEQ ID NO.1 in the sequence listing, and 30ng of the nucleotide sequence described in SEQ ID NO.2, 1 times of Mg 2+ PCR buffer solution, dNTP 2mmol, Taq DNA polymerase 1 unit, add sterile double distilled water to 10μl; put the special thin-walled tube for PCR amplification into the PCR instrument for amplification, the amplification conditions are: 94°C pre-denaturation for 180 seconds ; Denaturation at 94°C for 30 seconds, annealing at 55°C for 30 seconds, extension at 72°C for 60 seconds, 30 cycles; extension at 72°C for 7 minutes, the amplification is complete;

[0026] (3) Gel electrophore...

Embodiment 3

[0029] The method for detecting the purity of Jinyou No. 35 cucumber hybrid seeds comprises the steps:

[0030] (1) Genomic DNA was extracted from cucumber hybrid seeds of Jinyou 35;

[0031] (2) Carry out PCR amplification: Add Jinyou No. 35 cucumber hybrid seed genomic DNA 15ng in the special thin-walled tube for PCR amplification, then add 20ng of the nucleotide sequence described in SEQ ID NO.1 in the sequence listing, and the sequence listing 20ng of the nucleotide sequence described in SEQ ID NO.2, 1 times containing Mg 2+ PCR buffer, dNTP 2mmol, Taq DNA polymerase 0.8 unit, add sterile double distilled water to 10μl; put the thin-walled tube dedicated for PCR amplification into the PCR machine for amplification, the amplification conditions are: 94°C pre-denaturation for 180 seconds ; Denaturation at 94°C for 30 seconds, annealing at 55°C for 30 seconds, extension at 72°C for 60 seconds, 30 cycles; extension at 72°C for 7 minutes, the amplification is complete;

[003...

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Abstract

The invention relates to a primer sequence for detecting the purity of hybrid seeds of Jinyou No.35 cucumbers and a detection method. The primer sequence consists of an upstream primer and a downstream primer, wherein the upstream primer is a nucleotide sequence shown in SEQ ID NO.1 in a sequence table; and the downstream primer is a nucleotide sequence shown in SEQ ID NO.2 in the sequence table. The detection method has the characteristics of quickness, simpleness and convenience, stability, reliability, low cost and the like, and is not affected by environmental conditions. The identification on the purity of one batch of hybrid seeds of the Jinyou No.35 cucumbers can be finished within 5 to 6 hours. A large amount of manpower and soil fertility is saved, and identification results are quick and accurate. The primer sequence and the detection method have high application value on the identification of the purity of cucumber seeds.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a primer sequence and a detection method for detecting the purity of Jinyou 35 cucumber hybrid seeds. Background technique [0002] Seed purity identification is a key link to ensure seed quality. With the acceleration of the breeding process, the number of new cucumber varieties continues to increase, and the amount of materials that need to be identified for variety purity will increase. In the past, the conventional purity identification was carried out in the field. During the fruiting period, experts went to the field to observe the characteristics and consistency of the varieties. There was a long cycle, a heavy workload, and it was easily affected by environmental factors, and the phenotypic characteristics would have a certain degree of deviation. Problems such as these may affect the accuracy of variety purity identification and directly affect the promotion of improved var...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
Inventor 张桂华杜胜利王全崔兴华李鹏宇李加旺韩毅科魏爱民刘楠
Owner TIANJIN RES INST OF VEGETABLE
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