A kind of dna extraction method of horny layer of animal horn medicinal material

An extraction method and animal horn technology are applied in the field of identification of traditional Chinese medicinal materials, and can solve the problems of inability to perform PCR on horn medicinal materials, incomplete keratinocyte lysis, low DNA template concentration, etc., and achieve the effects of improving the incomplete lysis and the lysis effect.

Active Publication Date: 2020-10-16
SHANXI PROVINCE CHINESE MEDICINE RES INST
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Problems solved by technology

[0003] In order to solve the problems of incomplete keratinocyte lysis in the existing method for extracting DNA from animal horn medicinal materials, the concentration of the extracted DNA template is low, and PCR cannot be performed on horn medicinal materials with severe DNA degradation, the present invention provides an animal horn medicinal material cutin layer DNA extraction method

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  • A kind of dna extraction method of horny layer of animal horn medicinal material
  • A kind of dna extraction method of horny layer of animal horn medicinal material
  • A kind of dna extraction method of horny layer of animal horn medicinal material

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Embodiment 1

[0016] Example 1: Collect all horn animal medicinal materials and folk collection samples (including horns and horns) in Hebei Anguo medicinal materials market and Anhui Bozhou medicinal materials market, and study the DNA extraction method of horn medicinal materials (cuticle), so that the extracted The DNA quality can meet the requirements of conventional PCR reactions.

[0017] 1. Materials and Instruments

[0018] The collected sample information is shown in Table 1. For sample photos, see figure 1 . only by virtue of figure 1 In the photo, the species source information of the sample cannot be identified by traditional identification methods, and for samples with a small amount of DNA template, it is difficult to meet the requirements of PCR sequencing by conventional DNA extraction, so it is also impossible to identify the species source of the sample. Observation is easy to confuse the sample species, and misjudgment and misjudgment occur, which eventually lead to m...

experiment example

[0047] Experimental example: using the old croissant horn sample that has been preserved for more than 10 years, it is processed according to the conventional kit treatment method and the method of the present invention respectively. The conventional kit treatment method does not include the (3) step, and the method of the present invention includes the (3) step. Blood / cell / tissue genomic DNA extraction kit (spin column type) from Tiangen Biochemical Technology Co., Ltd., catalog number: DP304 (including Proteinase K, buffer GA, buffer GB, buffer GD, washing solution PW, adsorption column CB3). See Table 6 for sample information.

[0048] Table 6: Information on samples stored for more than 10 years

[0049] Numbering sample name Sample storage time place of origin 1 Antelope Horns (Size) 15 years siberia 2 Antelope horn block 13 years Russia 3 Tibetan antelope horns (height) 12 years Xinjiang

[0050] The specific steps are:...

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Abstract

The invention belongs to the technical field of identification of traditional Chinese medicinal materials. In order to solve the problems of existing methods for extracting DNA from animal horn medicinal materials, such as incomplete keratinocyte lysis, low concentration of extracted DNA templates, and the inability to perform PCR on horn medicinal materials with severe DNA degradation, a method is provided. DNA extraction method from the cuticle of animal horn medicinal materials. Sample pre-processing, sampling, DNA extraction, the extracted DNA samples are subjected to PCR amplification, and low-concentration samples are subjected to secondary amplification. It was determined that the sampling volume was 25mg, the DTT dosage was 20µl, the keratinocytes were completely lysed, and the DNA quality could meet the PCR requirements. Establish a DNA extraction method for the cuticle of horned animal medicinal materials so that the quality of the extracted DNA can meet the requirements of conventional PCR. The established DNA extraction method can extract DNA completely, can be operated in a standardized manner, and can be applied to the extraction of a variety of animal horns.

Description

technical field [0001] The invention belongs to the technical field of identification of traditional Chinese medicinal materials, and in particular relates to a method for extracting DNA from cuticles of animal horn medicinal materials. Background technique [0002] In Chinese medicine, some animal horns have significant clinical curative effects. For example, antelope horns have the effects of calming the liver and quenching wind, clearing the liver and improving eyesight, dispersing blood and detoxifying; buffalo horns have the effects of clearing heat and cooling blood, detoxifying, and calming shock. Due to the limited resources of horn animal medicinal materials, high prices, and many easily mixed products, traditional identification methods mainly rely on experienced identification experts. Generally, quality inspection personnel for traditional Chinese medicinal materials lack experience in character identification, and it is difficult to ensure the effectiveness of m...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/10C12Q1/6806
CPCC12N15/1003C12Q1/6806C12Q2531/113Y02A50/30
Inventor 张小慧刘霞杨方王长彪王世伟宋美卿
Owner SHANXI PROVINCE CHINESE MEDICINE RES INST
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