Method, kit and application for rapidly extracting genomic dna from hazelnut oil

A technology of toad oil and genome, which is applied in the field of genome DNA extraction of Chinese medicinal materials, can solve the problems of inability to carry out DNA extraction steps of toad oil medicinal materials, unfavorable DNA barcode identification, etc., to achieve efficient purification, fast and effective extraction, and fast release effect

Active Publication Date: 2019-05-10
哈尔滨市食品药品检验检测中心 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, the Chinese Pharmacopoeia animal medicine toad oil sample is not a fresh dry material sample, rich in mucus, colloid and hemicelluloses and other substances. The toad oil medicinal material is highly swollen after the solution, and the expansion degree is as high as more than 55 times. After centrifugation in a water bath, there is no supernatant or very little supernatant, and the subsequent DNA extraction steps of the toad oil medicinal material cannot be performed, which is not conducive to subsequent DNA barcode identification and other work.

Method used

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  • Method, kit and application for rapidly extracting genomic dna from hazelnut oil

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Example 1 Dried toad oil genome DNA extraction method

[0026] Establish a standard method for rapidly extracting the genomic DNA of the medicinal material of Haba oil, the specific extraction method is as follows:

[0027] 1. Use Chelex-100 and proteinase K as the lysate to lyse the dried toad oil sample.

[0028] 2. Use a grinding pestle to grind repeatedly until there are no obvious particles, and mix well.

[0029] 3. Keep warm in a water bath to make proteinase K play a role in digestion.

[0030] 4. Keep warm under boiling conditions to lyse the cells, and at the same time, under the chelating effect of Chelex-100 on non-nucleic acid organic matter, prevent the water absorption and swelling of the dried toad oil sample.

[0031] 5. Centrifuge the supernatant and use ddH 2 O replenish the volume of the supernatant to prevent the supernatant from being too small to better complete the subsequent extraction steps.

[0032] 6. Add impurity removal solution phenol / ...

Embodiment 2

[0042] Example 2 Large sample verification

[0043] In order to verify the general effectiveness of the dried toad oil genomic DNA extraction method, reagent composition and concentration determined in Example 1, various samples of toad oil were used for verification. Specifically:

[0044] 1) Realize the rapid DNA extraction of toad oil medicinal materials from different origins (Heilongjiang Province, Jilin Province and Liaoning Province) and the standard control medicinal materials of the Chinese Institute for the Control of Pharmaceutical and Biological Products.

[0045] 2) Realize the rapid DNA extraction of toad oil medicinal materials with different commercial specifications (line oil and block oil).

[0046] 1. Take about 5 mg of toad oil medicinal material, add lysate containing 400 μl 5% Chelex-100 and 4 μl proteinase K (20 mg / mL).

[0047] Among them, the dried toad oil medicinal materials used are:

[0048] Samples collected in Heilongjiang: C1, C2, and C3; C1,...

Embodiment 3

[0067] Example 3 Kit for Rapid Extraction of Toad Oil Genomic DNA

[0068] The kit includes at least the following components:

[0069] 1. Lysis solution: containing Chelex-100 and proteinase K, the concentration of Chelex-100 is 5%-10%, the concentration of proteinase K is 10-20mg / mL;

[0070] 2. Impurity removal solution: phenol / chloroform / isoamyl alcohol (25:24:1), used to remove adipose tissue and protein in toad oil samples.

[0071] In addition, the kit can also include reagents required for routine DNA extraction such as TE. A grinding pestle can also be included if desired.

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Abstract

The invention discloses a method for quickly extracting genome DNA from forest frog's oviduct, comprising the steps: (1) taking a forest frog's oviduct sample, adding Chelex-100 and protease K to the forest frog's oviduct sample, carrying out water bath heat insulation first, then terminating at high temperature, carrying out cell cracking, and centrifuging to obtain first supernatant, wherein the Chelex-100 is used for chelating non-nucleic acid organic matters in the forest frog's oviduct sample and inducing protein denaturation to facilitate the protease K to carry out enzymolysis, and the protease K is used for digesting protein in the forest frog's oviduct sample; (2) adding phenol / chloroform / isoamyl alcohol to the first supernatant for removing residual lipoid and protein; and (3) precipitating DNA by using isopropanol or absolute ethyl alcohol. The invention also provides a kit for quickly extracting genome DNA from the forest frog's oviduct, and application of a DNA bar code technology in identification of the forest frog's oviduct. The extraction method disclosed by the invention can realize quick and effective extraction of the genome DNA from dried forest frog's oviduct medicinal materials, and provides a basis for wider application of the DNA bar code technology.

Description

technical field [0001] The invention relates to the field of genomic DNA extraction of traditional Chinese medicinal materials, in particular to a method, a kit and an application thereof for rapidly extracting the genomic DNA of toad oil. Background technique [0002] The identification of traditional Chinese medicine is the premise and basis for studying the variety and quality of traditional Chinese medicine, formulating the standard of traditional Chinese medicine, and finding and expanding the source of medicine. The four traditional identification methods of traditional Chinese medicine are base identification, character identification, microscopic identification and physical and chemical identification. The theoretical basis of traditional identification methods is based on the analysis of the traits of taxa. These traits are phenotypes closely related to the growth and development stages of species and the environment, and are easily affected by subjective and object...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/10C12Q1/6806
CPCC12N15/1003C12Q1/6806C12Q2521/537C12Q2523/113
Inventor 唐先明石林春刘景波刘建辉宋经元刘金欣王洋顾佳王福宾孙志国
Owner 哈尔滨市食品药品检验检测中心
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