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A kind of asarum molecular ID card and its application

A technology of Asarum and DNA molecules, applied in the field of molecular biology, can solve problems such as inability to accurately identify species, affecting the accuracy of results, contaminating sequences, etc.

Active Publication Date: 2022-06-28
INST OF MEDICINAL PLANT DEV CHINESE ACADEMY OF MEDICAL SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] However, there are many species in the genus Asarum, and it is difficult to identify all species of the genus with traditional morphological methods; on the one hand, the current conventional DNA barcoding method cannot accurately identify each species in the genus, and for mixed samples with severe DNA degradation, Such as Chinese patent medicines or food mixtures, etc., the identification efficiency is low; bridge PCR and splicing in the process of high-throughput sequencing based on PCR products will cause certain contaminated sequences, which will affect the accuracy of the results

Method used

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  • A kind of asarum molecular ID card and its application
  • A kind of asarum molecular ID card and its application
  • A kind of asarum molecular ID card and its application

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Embodiment 1

[0036] Embodiment 1 The identification method of Asarum

[0037] 1) Download the full sequence of Asarum internal transcribed spacer (ITS) from NCBI, and obtain multiple candidate sequences by screening the sequence conserved regions of Asarum in ITS. Blast different candidate sequences in NCBI respectively, and screen out a conserved and specific sequence located in the ITS2 sequence region of Asarum; further cut out fragments of different lengths in the sequence region and perform Blast again to determine the As the best sequence fragment of Asarum molecular ID (Table 1, statistics of BLAST results of fragments located at different positions and with different lengths obtained by various interception methods). The alignment results showed that when the sequence fragment was longer, the sequence conservation within the genus Asarum decreased, and a single sequence could not cover all species in the genus ( figure 1 ); and when the sequence is too short, its specificity will ...

Embodiment 2

[0058] Example 2 Identification of Asarum samples

[0059] 1) After grinding Asarum sp. samples in liquid nitrogen, total DNA was extracted with a plant genomic DNA extraction kit from Tiangen Biochemical Technology (Beijing) Co., Ltd.

[0060] 2) High-throughput sequencing of the mixed DNA samples directly on the Illumina platform.

[0061] 3) Extract the molecular ID sequence of Asarum from the original data obtained by high-throughput sequencing, and the results are as follows Image 6 .

[0062] 4) The results show that the molecular ID sequence SEQ ID No. 1 can also be accurately detected in the samples of broken Asarum. It is proved that the presence of Asarum in a sample can be determined by directly detecting the presence of reads containing molecular IDs in high-throughput raw sequencing data.

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Abstract

The invention relates to the technical field of molecular biology, in particular to an asarum molecular ID card and its application. The Asarum molecular ID card contains the nucleotide sequence shown in SEQ ID NO.1 or SEQ ID NO.1‑5. The present invention also provides a method for identifying the genus Asarum, which includes determining whether the nucleotide sequence shown in SEQ ID NO.1 or SEQ ID NO.1-5 is contained in the mixed sample by detecting whether the high-throughput original sequencing data contains Presence of Asarum species. The molecular ID card and method provided by the invention can be used to accurately identify Asarum. The method provided by the invention has wider applicability and can detect whether all samples from which DNA can be extracted contain Asarum species. Therefore, rapid and accurate identification of Asarum original plants, medicinal materials, seeds, seedlings, powders and food mixtures can be realized.

Description

technical field [0001] The invention relates to the technical field of molecular biology, in particular to a molecular ID card belonging to the genus Asarum and its application. Background technique [0002] Asarum is a perennial herb, and there are about 90 species in the world. There are 30 species, 4 varieties and 1 variant distributed in my country. They are distributed all over the north and south, and the provinces and regions south of the Yangtze River Basin are the most. Asarum has the functions of relieving the exterior and dispelling cold, dispelling wind and relieving pain, and clearing the orifices, warming the lungs and transforming Yin. . As a commonly used Chinese medicinal material, Asarum has a long history of application, but the plants of this genus contain aristolochic acids (AAs), which need to be processed before being used as medicine. As one of the strongest carcinogens in the Carcinogenicity Database (CPDB), AAs has accumulated toxicity in the huma...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895
Inventor 韩建萍白璇姣王刚宋经元
Owner INST OF MEDICINAL PLANT DEV CHINESE ACADEMY OF MEDICAL SCI