Triplex PCR method for identifying purity of sweet melon seed

A technology for the purity and purity of hybrids, which is applied in the direction of biochemical equipment and methods, and the determination/inspection of microorganisms, can solve the problems that have not been seen in the identification of the purity of melon seeds by multiple PCR, and achieve shortened working hours, high accuracy, and reduced The effect of consumption

Active Publication Date: 2018-08-21
HENAN AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
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Problems solved by technology

But generally speaking, in the prior art, there is no good related report on the identification of the purity of melon seeds by using multiplex PCR technology.

Method used

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  • Triplex PCR method for identifying purity of sweet melon seed
  • Triplex PCR method for identifying purity of sweet melon seed
  • Triplex PCR method for identifying purity of sweet melon seed

Examples

Experimental program
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Effect test

Embodiment 1

[0046] (1) Extraction and preparation of DNA templates from materials to be tested

[0047] Generally speaking, when extracting muskmelon genomic DNA, it is convenient and easy to operate with young leaves at the seedling stage. The specific process is briefly introduced as follows.

[0048] The melon seeds to be tested were soaked in warm soup to accelerate germination. Sow 10 plants of DHL92 and TopMarker materials in seedling trays respectively as control group 1 and control group 2. The fresh-keeping box was used as the experimental group. After sowing, three layers of absorbent paper were covered on the top of the seeds and watered, and the seedlings were raised in the light incubator, and the DNA of the young leaves was extracted when about two leaves were one heart.

[0049] Genomic DNA was extracted for each group of biological material samples, and the specific operations were as follows:

[0050] Add 2 steel balls into a 2ml centrifuge tube, tear off about 1.5cm le...

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Abstract

The invention belongs to the technical field of sweet melon cultivation and in particular relates to a triplex PCR method for identifying the purity of a sweet melon seed. The method is applied to hybrid seed purity identification and specifically comprises the following steps: extracting DNA of a material to be detected, performing triplex PCR amplification and electrophoresis detection, performing analysis and evaluation, and the like. By adopting the triplex PCR method for identifying the purity of the sweet melon seed, a plurality of sites can be simultaneously identified in a same reaction, so that the working time can be shortened, and in addition, the consumption of test reagent consumables can be reduced in times. When the purity of the sweet melon seed is identified by using the method, the defect that a conventional single PCR method is slow in velocity in seed purity identification can be overcome, the method has the characteristics of being short in time, good in specificity, high in accuracy, low in cost, wide in applicability, and the like, the purity of a sweet melon seed material can be simply, rapidly and conveniently obtained, and thus subsequent germplasm resource analysis and seed breeding can be relatively specifically carried out.

Description

technical field [0001] The invention belongs to the technical field of melon cultivation, and specifically relates to a patent application for a triple PCR identification method for identifying the purity of melon seeds. Background technique [0002] melon( Cucumis melo L) for Cucurbitaceae ( Cucurbitaceae ) Melon ( Cucumis ) annual herbaceous vine originating from India and its adjoining areas. my country is a major producer of melons. In 2015, the cultivated area of ​​melons in my country reached 460,900 hectares, with an output of 15.271 million tons. [0003] During the cultivation of muskmelons, the purity of varieties is an important prerequisite to ensure the consistency of muskmelon yield and management measures. Therefore, testing the purity of varieties is an important prerequisite for related cultivation, breeding and other work. In the prior art, the identification of seed purity is a relatively heavy task, especially the rich variation phenotypes of melon...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6858C12Q1/6895
CPCC12Q1/6858C12Q1/6895C12Q2600/156C12Q2600/16C12Q2531/113C12Q2537/143C12Q2565/125
Inventor 杨路明马长生刘东明胡倩梅朱华玉胡建斌
Owner HENAN AGRICULTURAL UNIVERSITY
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