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Asarum molecular identity card and application thereof

A molecular ID card and Asarum technology, applied in the field of molecular biology, can solve the problems of low identification efficiency, affecting the accuracy of results, contaminating sequences, etc.

Active Publication Date: 2021-06-18
INST OF MEDICINAL PLANT DEV CHINESE ACADEMY OF MEDICAL SCI
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  • Summary
  • 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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  • Asarum molecular identity card and application thereof
  • Asarum molecular identity card and application thereof
  • Asarum molecular identity card and application thereof

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Embodiment 1

[0036] The identification method of embodiment 1 Asarum

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

Embodiment 2

[0058] The identification of embodiment 2 Asarum samples

[0059] 1) After grinding Asarum sp. in liquid nitrogen, the total DNA was extracted with the Plant Genomic DNA Extraction Kit of Tiangen Biochemical Technology (Beijing) Co., Ltd.

[0060] 2) The mixed DNA samples were directly subjected to high-throughput sequencing on the Illumina platform.

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

[0062] 4) The results show that the molecular identity card sequence SEQ ID No.1 can also be accurately detected in the samples of Asarum spp. It proves that it is possible to judge whether there is Asarum in a sample by directly detecting whether there are reads containing molecular ID cards in the 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 identity card and application thereof. The asarum molecular identity card contains a nucleotide sequence as shown in SEQ ID NO.1 or SEQ ID NO.1-5. The invention further provides an asarum identification method. The asarum identification method comprises the step of judging whether the asarum species exist in a mixed sample or not by detecting whether the nucleotide sequence as shown in SEQ ID NO.1 or SEQ ID NO.1-5 is contained in high-throughput original sequencing data. By utilizing the molecular identity card and the method, the asarum can be accurately identified, and the method is wider in applicability and can be used for detecting whether all samples from which DNA can be extracted contain the asarum species or not. Therefore, asarum original plants, medicinal materials, seeds, seedlings, powder and food mixtures can be rapidly and accurately identified.

Description

technical field [0001] The invention relates to the technical field of molecular biology, in particular to an asarum molecular ID card and its application. Background technique [0002] Asarum is a perennial herb, and there are about 90 species in the world. In my country, there are 30 species, 4 varieties, and 1 variant. They are distributed in all parts of the north and south, and the provinces south of the Yangtze River Basin are the most. Asarum has the effects of relieving the exterior and dispersing cold, dispelling wind and relieving pain, clearing the orifices, warming the lungs and transforming the drink, and is often used to treat colds, headaches, toothaches, nasal congestion, runny nose, nasal congestion, nasal sinusitis, rheumatism arthralgia, phlegm retention, asthma and coughing . As a commonly used traditional Chinese medicinal material, Asarum has a long history of application, but this genus contains aristolochic acids (AAs), which need to be processed bef...

Claims

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Application Information

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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