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SNP marker of medicinal plant murraya paniculata as well as identifying method and application thereof

A technology for medicinal plants and gulira, which is applied in the biological field to achieve the effects of simple identification process, stability and reliability, and wide applicability.

Inactive Publication Date: 2016-08-03
CHINA RESOURCES SANJIU MEDICAL & PHARMA +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] However, in the prior art, an accurate and rapid method for identifying medicinal Murata sativa by using DNA barcodes or SNP markers has not been proposed

Method used

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  • SNP marker of medicinal plant murraya paniculata as well as identifying method and application thereof
  • SNP marker of medicinal plant murraya paniculata as well as identifying method and application thereof
  • SNP marker of medicinal plant murraya paniculata as well as identifying method and application thereof

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Experimental program
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Effect test

Embodiment 1

[0049] The rapid molecular identification method of the medicinal plant Murata chinensis described in the present embodiment comprises the following steps:

[0050] S1. Extract the total DNA of each of the samples of Murata to be tested according to conventional methods, and the extraction steps are as follows: take the leaves (about 30 mg) of the samples of Murata to be tested respectively, and grind them on a MM400 ball mill, Then use the Plant Genomic DNA Extraction Kit (TiangenBiotechCo., China) to extract the total genomic DNA for subsequent use;

[0051] S2. Using the total genomic DNA obtained in step S1 as a template, design the following primers:

[0052] ITS2F:5'-ATGCGATACTTGGTGTGAAT-3',

[0053] ITS3R:5'-GACGCTTCTCCAGACTACAAT-3', said primers are shown in SEQIDNo.2 and SEQIDNo.3, carry out PCR amplification to described total genomic DNA, and described PCR amplification reaction system comprises:

[0054] DNA template, 50ng, 2μL;

[0055] Forward and reverse prim...

Embodiment 2

[0061] The rapid molecular identification method of the medicinal plant Murata chinensis described in the present embodiment comprises the following steps:

[0062] S1. According to the conventional method, extract the total DNA of the medicinal Murata to be tested and the samples of the closely related species of Murata. The extraction steps are as follows: take about 30 mg of the leaves of the samples to be tested respectively, and conduct them on the MM400 ball mill. Grind, then extract the total genomic DNA with a plant genomic DNA extraction kit (TiangenBiotechCo., China), and set aside;

[0063] S2. Using the total genomic DNA obtained in step S1 as a template, design the following primers:

[0064] ITS2F:5'-ATGCGATACTTGGTGTGAAT-3',

[0065] ITS3R: 5'-GACGCTTCTCCAGACTACAAT-3', performing PCR amplification on the total genomic DNA, the PCR amplification reaction system comprising:

[0066] DNA template, 50ng, 2μL;

[0067] Forward and reverse primers, 2.5 μM, 1 μL each...

Embodiment 3

[0073] The rapid molecular identification method of the medicinal plant Murata chinensis described in the present embodiment comprises the following steps:

[0074] S1. Extract the total DNA of each sample of the medicinal Murata to be tested according to a conventional method. The extraction steps are as follows: take 1 seed of the sample of Murata to be tested, grind it on a MM400 ball mill, and then use Plant Genomic DNA Extraction Kit (TiangenBiotechCo., China) was used to extract the total genomic DNA and set aside;

[0075] S2. Using the total genomic DNA obtained in step S1 as a template, design the following primers:

[0076] ITS2F:5'-ATGCGATACTTGGTGTGAAT-3',

[0077] ITS3R:5'-GACGCTTCTCCAGACTACAAT-3', said primers are shown in SEQIDNo.2 and SEQIDNo.3, carry out PCR amplification to described total genomic DNA, and described PCR amplification reaction system comprises:

[0078] DNA template, 40ng, 2μL;

[0079] Forward and reverse primers, 2.5 μM, 1 μL each;

[008...

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Abstract

The invention provides an SNP marker of a medicinal plant murraya paniculata. In the SNP marker, the nucleotide on the 75th site from the 5' terminal in the sequence shown by SEQ ID No.1 is T, the nucleotide on the 109th site is T and / or the nucleotide on the 173rd site is A; the sequence shown by SEQ ID No.1 is the ITS2 intergenic region sequence of murraya paniculata. By adopting the SNP marker, the medicinal murraya paniculata can be quickly and accurately identified; the SNP marker has wide applicability and can guarantee the stability and reliability of the identification result from the source; and compared with traditional morphological identification method, the SNP marker of the medicinal plant murraya paniculata can realize quick and accurate identification of the medicinal murraya paniculata.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a SNP marker of a medicinal plant Murata japonica and its identification method and application. Background technique [0002] Murraya exotica L. is a plant of the genus Murraya Koenigex Linnaeus in the family Rutaceae, and is produced in the south of five provinces, including Taiwan, Fujian, Guangdong, Hainan and Guangxi. The chemical components contained in the roots, stems, and leaves of Murrayapaniculata (L.) Jack. are similar to those of Murrayapaniculata (L.) Jack. Jiulixiang has the effects of promoting qi and relieving pain, promoting blood circulation and dissipating blood stasis, expelling wind and dampness, anesthesia and convulsions, etc. It is clinically used to treat stomach pain, rheumatic arthralgia, toothache, tumbling pain and insect bites. Jiulixiang M.exoticaL. is full of treasures. In addition to its own medicinal value, it is often used as an orname...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6895C12Q1/6858C12Q2600/156C12Q2531/113
Inventor 姚辉周红马双姣陈士林宋经元陈贝贝邢建永杨锐培马鹏岗
Owner CHINA RESOURCES SANJIU MEDICAL & PHARMA
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