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Primer sequence and detection method for detecting purity of Jin superior NO.36 cucumber hybridized seeds

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

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

AI Technical Summary

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 and detection method for detecting purity of Jin superior NO.36 cucumber hybridized seeds
  • Primer sequence and detection method for detecting purity of Jin superior NO.36 cucumber hybridized seeds

Examples

Experimental program
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Embodiment 1

[0017] The method for detecting the purity of Jinyou No. 36 cucumber hybrid seeds comprises the steps:

[0018] (1) Extracting the genomic DNA of the root tip of Jinyou 36 cucumber hybrid seeds 36 hours after germination;

[0019] (2) Carry out PCR amplification: Add Jinyou No. 36 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 minute...

Embodiment 2

[0023] The method for detecting the purity of Jinyou No. 36 cucumber hybrid seeds comprises the steps:

[0024] (1) Genomic DNA was extracted from cucumber hybrid seeds of Jinyou 36;

[0025] (2) Perform PCR amplification: add 20ng of Jinyou 36 cucumber hybrid seed genomic DNA in a 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 electrophoresi...

Embodiment 3

[0029] The method for detecting the purity of Jinyou No. 36 cucumber hybrid seeds comprises the steps:

[0030] (1) Genomic DNA was extracted from cucumber hybrid seeds of Jinyou 36;

[0031] (2) Carry out PCR amplification: Add Jinyou No. 36 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 primer sequence and detection method for detecting purity of Jin superior NO.36 hybridized cucumber seeds, wherein the primer sequence consists of a forward primer and a reverse primer, the forward primer is a nucleotide sequence shown in SEQ ID NO.1 in a sequence table, and the reverse primer is a nucleotide sequence shown in SEQ ID NO.2 in the sequence table. The primer sequence and detection method provided by the invention have the advantages of being fast, simple, convenient, stable and reliable, having low cost and being not affected by environment conditions, and the like, can be used for carrying out complete identification of one batch only for 5-6 hours on the purity of the Jin superior NO.36 hybridized cucumber seeds, thus saving a great deal of manpower and land capacity, being fast and accurate in identification result, and having great application value in the purity identification of the 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 36 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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