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Maize variety Shandan 609 purity detection InDel molecular marker kit

A technology of purity detection and molecular markers, which is applied to the determination/inspection of microorganisms, biochemical equipment and methods, etc., and can solve the problems that cannot meet the requirements of corn seed purity detection

Inactive Publication Date: 2019-08-02
ANHUI SCI & TECH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the practice of traditional agricultural production, the identification of corn seed purity is mainly based on grain morphology identification, seedling identification and field planting identification, supplemented by biochemical isoenzyme and protein electrophoresis technology, but with the homogeneity of corn germplasm resources The above two methods have been unable to meet the detection requirements of corn seed purity.

Method used

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  • Maize variety Shandan 609 purity detection InDel molecular marker kit
  • Maize variety Shandan 609 purity detection InDel molecular marker kit
  • Maize variety Shandan 609 purity detection InDel molecular marker kit

Examples

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Effect test

Embodiment 1

[0036] In this embodiment, the purity detection InDel molecular marker kit of the corn variety Shandan 609 was used to detect the purity of the corn variety Shandan 609 sample A, and the operation process was as follows:

[0037] (1) A150 seeds of the corn variety Shandan 609 sample were randomly selected, and DNA was extracted by the CTAB method;

[0038] (2) PCR amplification, using sequences 1 and 2 as InDel-specific primers, prepare a 15 μL PCR reaction system;

[0039] (3) The total volume of the PCR reaction system is 15 μL. Including 10.2 μL ultrapure water, 1.5 μL 10-fold PCR buffer, 1.2 μL dNTPs (2.5 mmol / L), 1.0 μL InDel primer (0.25 μmol / L), 0.15 μL Taq DNA polymerase (2.5 U / μL), 1 μL DNA, mixed uniform;

[0040] (4) The reaction program is pre-denaturation at 94°C for 5 minutes; denaturation at 94°C for 1 minute, annealing at 55°C for 40 seconds, extension at 72°C for 40 seconds, and 35 cycles; extension at 72°C for 10 minutes; storage at 4°C;

[0041] (5) Elect...

Embodiment 2

[0047] In this embodiment, the purity detection InDel molecular marker kit of the corn variety Shandan 609 was used to detect the purity of the corn variety Shandan 609 sample B, and the operation process was as follows:

[0048] (1) Take B200 seeds of the corn variety Shandan 609 sample at random, and take the CTAB method to carry out DNA extraction;

[0049] (2) PCR amplification using sequences 3 and 4 as InDel-specific primers to prepare a 15 μL PCR reaction system;

[0050](3) The total volume of the PCR reaction system is 15 μL, including 10.2 μL ultrapure water, 1.5 μL 10X PCR buffer, 1.2 μL dNTPs (2.5 mmol / L), 1.0 μL InDel primer (0.25 μmol / L), 0.15 μL Taq DNA polymerase (2.5U / μL), 1μL DNA, mix well;

[0051] (4) The reaction program is pre-denaturation at 94°C for 5 minutes; denaturation at 94°C for 1 minute, annealing at 55°C for 40 seconds, extension at 72°C for 40 seconds, and 35 cycles; extension at 72°C for 10 minutes; storage at 4°C;

[0052] (5) Electrophores...

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Abstract

The invention discloses a maize variety Shandan 609 purity detection InDel molecular marker kit, and relates to a molecular marker detection kit, which utilizes the male and female parental complementary specific InDel primer to detect the purity of the maize variety Shandan 609, and utilizes the genome DNAs of the maize variety Shandan 609 and male and female parents as a template. By screening the whole genome InDel primers, two pairs of parental complementary primers which are complementary, highly specific, highly reproducible and stable in result are obtained. A kit for the purity detection of the maize variety Shandan 609 is developed by using the two specific primers and reagents such as Taq enzyme. The kit can quickly detect the purity of maize seeds, provide scientific and technical support for the market supervision of maize varieties, and is of great significance for improving the quality of crop seeds and ensuring food safety production.

Description

technical field [0001] The invention belongs to a molecular marker kit, in particular to an InDel molecular marker kit for purity detection of a corn variety Shandan 609. Background technique [0002] InDel insertion and deletion markers refer to the difference in the whole genome of the two parents. Compared with the other parent, one parent has a certain number of nucleotide insertions or deletions in the genome, according to the insertion and deletion sites in the genome , design some PCR primers to amplify these indel sites. As a high-throughput molecular marker, InDel has the advantages of various genetic markers. The development of traditional molecular markers is generally based on only one sequence, while the development of InDel markers is completely based on sequence differences, so there are fewer polymorphic sites during the development process, and the probability of mutation is extremely low, so InDel markers are more accurate and stable in mutation, avoiding ...

Claims

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Application Information

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IPC IPC(8): C12Q1/6895
CPCC12Q1/6895C12Q2600/156
Inventor 王歆
Owner ANHUI SCI & TECH UNIV
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