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Extracting solution for extracting DNA from micro medicinal plant sample and extraction method

A technology of medicinal plants and extraction methods, applied in the field of DNA extraction, can solve the problems of large manpower, material resources and time, scarce samples, expensive kits, etc., to promote development and popularization, simple operation process, plant samples less effect

Pending Publication Date: 2019-12-17
LISHUI AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the kit is expensive and not suitable for batch sample operation, and it takes a lot of manpower, material resources and time to identify batch samples
In addition, the samples of some rare and endangered medicinal plants are extremely rare, and it is difficult to obtain sufficient samples for the extraction of genomic DNA. Currently, there are no reagents and methods suitable for batch extraction of rare samples on the market
Molecular identification method has become a major development trend in the identification of traditional Chinese medicine, but the above-mentioned problems seriously restrict the use and promotion of molecular identification method

Method used

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  • Extracting solution for extracting DNA from micro medicinal plant sample and extraction method
  • Extracting solution for extracting DNA from micro medicinal plant sample and extraction method
  • Extracting solution for extracting DNA from micro medicinal plant sample and extraction method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0068] The genomes of fresh leaves of various medicinal plants included in the "Chinese Pharmacopoeia" were extracted to amplify the DNA barcode core sequence ITS2, and the species were identified through the sequence information.

[0069] (1) For sample

[0070] A total of 16 fresh samples of medicinal plant leaves of different species included in the "Chinese Pharmacopoeia" were collected, as shown in Table 1.

[0071] Table 1 Sample information sheet

[0072]

[0073] Among them, Huachonglou is named "Aesculus chinensis" in "Chinese Pharmacopoeia"; palm leaf raspberry is named "East China raspberry" in "Chinese Pharmacopoeia".

[0074] (2) Genomic DNA extraction

[0075] The DNA of the test sample was extracted according to the DNA extraction method described above.

[0076] (3) PCR amplified DNA barcode sequence (ITS2)

[0077] All the tested samples could amplify a single bright band by PCR, such as figure 1 As shown, subsequent experiments can be carried out.

...

Embodiment 2

[0081] The leaf stem genomes of various medicinal plants collected in the "Chinese Pharmacopoeia" were extracted to amplify the DNA barcode core sequence ITS2, and the species were identified through the sequence information.

[0082] (1) Provide samples

[0083] A total of 16 dry samples of medicinal plant leaves of different species included in the "Chinese Pharmacopoeia" were collected, as shown in Table 2.

[0084] Table 2 Sample information table

[0085]

[0086] (2) Genomic DNA extraction

[0087] The DNA of the test sample was extracted according to the DNA extraction method described above.

[0088] (3) PCR amplified DNA barcode sequence (ITS2)

[0089] All the tested samples could amplify a single bright band by PCR, such as Figure 18 As shown, subsequent experiments can be carried out.

[0090] (4) Sequence PCR fragments, obtain sequence information, and identify species online through the Chinese herbal medicine DNA barcode identification system.

[0091]...

Embodiment 3

[0093] Extract the genomes of different parts fresh samples (tissues with the same medicinal parts) of various non-leaf medicinal plants in Example 1 and Example 2 to amplify the DNA barcode core sequence ITS2, and pass the sequence information Identify species.

[0094] (1) Provide samples

[0095] A total of 16 fresh samples of different parts of medicinal plants of different species included in the Chinese Pharmacopoeia were collected, see Table 3 for details.

[0096] Table 3 Sample Information Sheet

[0097]

[0098]

[0099] (2) Genomic DNA extraction

[0100] The DNA of the test sample was extracted according to the DNA extraction method described above.

[0101] (3) PCR amplification of DNA barcode sequence (ITS2).

[0102] All the tested samples could amplify a single bright band by PCR, such as Figure 19 As shown, subsequent experiments can be carried out.

[0103] (4) Sequence PCR fragments, obtain sequence information, and identify species online throu...

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Abstract

The invention discloses an extracting solution for extracting DNA from a micro medicinal plant sample and an extraction method. The extracting solution is prepared from the components: 10-20 mmol / L ofTris-HCl, 0.5-1 mmol / L of EDTA, 0.1-0.2 mmol / L of KCl and 5-10 mg / L of RNA enzyme. The extracting solution can be used for extracting genomic DNA of a micro sample from a variety of tissue such as leaves, stems, roots and flowers of various medicinal plants, and is wide in applicability; the extracted genomic DNA by the extraction method can be effectively used for PCR amplification of target fragments and DNA barcode molecular identification. Compared with the prior art, the extraction method has the advantages of wide applicability, small amount of required samples, no high-toxic and highlytoxic components, low cost, simple operation, speediness, easy grasp and the like, a new method is provided for genomic DNA extraction and molecular identification of a micro sample of a medicinal plant based on a DNA barcode, and the development and popularization of molecular identification technology of traditional Chinese medicine are promoted.

Description

technical field [0001] The invention relates to the technical field of DNA extraction, more specifically, it relates to an extraction solution and an extraction method for extracting a trace amount of medicinal plant sample DNA. Background technique [0002] There are a wide variety of Chinese herbal medicines, and their sources are complex, and multi-sources are common. As a result, in the process of production, distribution and clinical use of traditional Chinese medicines, there are common names and different names of medicines, same names and different names, unclear identification, substitution and mixed use, false ones, and substandard ones. Phenomena such as filling up have had a strong impact on the safety and effectiveness of traditional Chinese medicine in the process of clinical application. Accurate identification of medicinal materials is the premise and basis for ensuring the quality of traditional Chinese medicine and the safety, effectiveness and controllabil...

Claims

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Application Information

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IPC IPC(8): C12N15/10C12Q1/6806
CPCC12N15/1006C12Q1/6806
Inventor 吕群丹方洁程科军潘俊杰陈正道
Owner LISHUI AGRI SCI
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