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S.nitidum (Vahl) Pers. testing primer and testing method

A detection method, sorghum technology, applied in the direction of DNA / RNA fragments, recombinant DNA technology, etc., can solve the problems of increasing and difficult identification of false sorghum, and the long cycle of cell biology methods, so as to shorten the detection time and improve the detection sensitivity Effect

Inactive Publication Date: 2008-08-20
印丽萍
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Y.Sun et al. (1994) thought that sorghum light should be included in the Sorghum block group, while DeWet (1978), Guo (1996) and others suggested that it should be incorporated into Parasorghum. increased difficulty
[0003] In actual work, the appearance characteristics of weed seeds such as false sorghum mixed in agricultural products are often worn and deformed due to transportation. At the same time, due to the influence of environment, climate, cultivation conditions, and differences in seed maturity, the individual differences Morphological identification has increased the difficulty, and cell biology methods not only require a longer cycle, but also are difficult to identify based on the morphology of chromosomes

Method used

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  • S.nitidum (Vahl) Pers. testing primer and testing method
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  • S.nitidum (Vahl) Pers. testing primer and testing method

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Embodiment Construction

[0063] 1 seed DNA acquisition

[0064] The single seed is ground and pulverized, and extracted according to the following steps:

[0065] 1) Grind the seeds in liquid nitrogen, put them into a 1.5mL centrifuge tube, add 500μL TES, TES is composed of 100mM Tris (pH8.0), 10mM EDTA, 2% SDS;

[0066] 2) Add 7 μL of proteinase K and mix well, place in a 55°C water bath for 60-120 minutes, and mix well several times during the period;

[0067] 3) Adjust the salt concentration to 1.4M, add 1 / 10 volume of 10% cetyltrimethylammonium bromide, and place in a water bath at 65°C for 10 minutes;

[0068] 4) Add an equal volume of SEVGA and mix well, SEVGA is chloroform:isoamyl alcohol=24:1, not too vigorously, to ensure the integrity of DNA, incubate at 0°C for 30min;

[0069] 5) Centrifuge at 13,000r / min at 4°C for 10min, and transfer the supernatant to a 1.5mL centrifuge tube;

[0070] 6) Add 225 μL of 5M ammonium acetate and mix well, incubate at 0°C for more than 30 minutes, and cent...

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Abstract

The invention relates to a shining sorghum specific primer and a PCR detection method by utilizing the specific primer, which overcomes the difficulties in the methods that are related to the morphologic identification and the cell biology in the prior art and is applicable to the fields of port inspection and quarantine, agricultural production, plant protection, and so on. The PCR detection method includes the steps of 1) the obtainment of a seed DNA, 2) PCR amplification and 3) agarose gel electrophoresis.

Description

(1) Technical field [0001] The invention belongs to the technical field of agricultural plant inspection and quarantine, and in particular relates to a detection method for quickly and accurately detecting light sorghum using molecular biology detection technology and a detection primer used in the method, which is suitable for port inspection and quarantine, agricultural production, and plant protection. used in other fields. (2) Background technology [0002] Light sorghum (Sorghum.nitidum (Vahl) Pers) is an approximate species of pseudo-sorghum, one of the world's top ten malignant weeds. Light sorghum was first discovered in western India, and then spread to Southeast Asia, Indonesia, Australia and other countries. Sorghum glabra is perennial and produces rhizomes similar in morphology and chromosome size to Sorghum pseudosorghum. Y.Sun et al. (1994) thought that sorghum light should be included in the Sorghum block group, while DeWet (1978), Guo (1996) and others sugge...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
Inventor 印丽萍易建平王伟
Owner 印丽萍