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DNA extraction method of animal horn pharmaceutical cuticle

An extraction method and a technology of animal horns, which are applied in the field of identification of Chinese medicinal materials, can solve problems such as inability to perform PCR on horny medicinal materials, low concentration of DNA templates, and incomplete lysis of keratinocytes, so as to improve the effect of lysis and incomplete lysis

Active Publication Date: 2017-07-28
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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  • DNA extraction method of animal horn pharmaceutical cuticle
  • DNA extraction method of animal horn pharmaceutical cuticle
  • DNA extraction method of animal horn pharmaceutical cuticle

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Embodiment 1

[0016] Example 1: Collect all horny animal medicinal materials in Hebei Anguo medicinal material market and Anhui Bozhou medicinal material market and samples of folk collections (including horns and horns), study the DNA extraction method of horny medicinal materials (stratum corneum), and make 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. See the sample photos figure 1 . Only by figure 1 Photos. Traditional identification methods cannot identify the species source information of the sample. For samples with a small amount of DNA template, conventional DNA extraction is difficult to meet the requirements of PCR sequencing. Therefore, the species source of the sample cannot be identified. Observation is easy to confuse the sample species, and misjudgment or misjudgment occurs, which eventually leads to medication errors.

[0019] Table 1: Co...

experiment example

[0047] Experimental example: the old horn samples stored for more than 10 years were processed according to the conventional kit processing method and the method of the present invention. The conventional kit processing method does not include step (3), and the method of the present invention includes step (3). Using Tiangen Biochemical Technology Co., Ltd. blood / cell / tissue genomic DNA extraction kit (spin column type), catalog number: DP304 (including Proteinase K, buffer GA, buffer GB, buffer GD, rinse PW, adsorption Column CB3). The sample information is shown in Table 6.

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

[0049] Numbering sample name Sample storage time Origin 1 Antelope horns (length) 15 years Siberia 2 Antelope Horn Block 13 years Russia 3 Tibetan antelope horns (length) 12 years Xinjiang

[0050] The specific steps are:

[0051] (1) Sample pretreatment: Take the corner medicinal material samples in Table 6, first cut the cross-secti...

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Abstract

The invention belongs to the technical field of traditional Chinese medicine identification, is intended to solve the problems that the existing animal horn pharmaceutical DNA extraction method has poor keratinocyte lysis, extracted DNA template concentration is low and horn pharmaceutical materials with severe DNA degradation are unavailable for PCR (polymerase chain reaction), and provides a DNA extraction method of animal horn pharmaceutical cuticle, comprising: pretreating a sample, sampling, extracting DNA, subjecting the extracted DNA sample to PCR amplification, and subjecting the low-concentration sample to secondary amplification. The sampling quantity of 25 mg is determined, DTT (DL-Dithiothreitol) usage is 20 Mul, keratinocytes split completely, and DNA quality may meet the PCR requirements. The DNA extraction method of pharmaceutical cuticle of horny animals is established herein such that the quality of the extracted DNA meets the requirements of conventional PCR. The DNA extraction method established herein allows DNA to be fully extracted, is available for standard operation and is applicable to the extraction of various animal horns.

Description

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

Claims

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Application Information

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IPC IPC(8): C12N15/10C12Q1/68
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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