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A technology of kits and medicinal materials, applied in the field of rapid DNA extraction kits from medicinal materials, which can solve the problems of large differences in plant materials and difficulty in finding formulas for alkali lysing agents
Inactive Publication Date: 2015-05-20
INST OF CHINESE MATERIA MEDICA CHINA ACAD OF CHINESE MEDICAL SCI
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Problems solved by technology
However, due to the huge difference in plant materials, it is difficult to find a general formula of alkali lysin
Method used
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Embodiment 1
[0021] 1. Rapid extraction of DNA from snake medicinal materials
[0022] A rapid DNA extraction kit from medicinal materials, comprising an extraction buffer (solution A, reagent C), a neutralization buffer (solution B) and an instruction manual. Wherein solution A is 1% tritonX-100 aqueous solution, solution B is 0.1M Tris-HCl aqueous solution, reagent C is NaOH and PVP. Use this kit for rapid DNA extraction of snake medicinal materials, the steps are as follows:
[0023] (1) Dissolve reagent C (0.08gNaOH, 0.04gPVP) in solution A (4mL) to make DNA extraction buffer, and dissolve it completely before use.
[0024] (2) Take 25mg each of the dried medicinal materials of black snake, golden snake, jade brocade snake, and red chain snake snake, and put them into 2mL centrifuge tubes respectively. After numbering, use a ball mill to grind the medicinal materials into powder for about 5 minutes, and set aside .
[0025] (3) Add 200 μL of DNA extraction buffer to the 4 samples re...
Embodiment 2
[0032] 1. Rapid extraction of DNA from plant medicinal materials
[0033] (1) Dissolve reagent C (0.2g NaOH, 0.1g PVP) in 10mL solution A (1% tritonX-100 aqueous solution) to prepare DNA extraction buffer, completely dissolved before use.
[0034] (2) Take turmeric, acacia bark, angelica, honeysuckle, ginseng, plantain seed, windproof, patchouli, safflower, flavescens, magnolia, foreknowledge seed, salvia miltiorrhiza, corydalis, platycodon grandiflorum, fritillaria, peony root, Put 25mg each of the dried medicinal materials of Duckweed, Polygonum multiflorum, Chinese yam, and Codonopsis pilosula into 2mL centrifuge tubes. After numbering, use a ball mill to grind the medicinal materials into powder for about 5 minutes, and set aside.
[0035] (3) Add 200 μL of DNA extraction buffer to 21 samples respectively, and vortex to mix well.
[0036] (4) Add 800 μL of solution B (0.1 M Tris-HCl aqueous solution) to 21 samples respectively, vortex to mix well, centrifuge at 12000 rpm ...
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Abstract
The invention discloses a rapid DNA extraction kit of a medicinal material, belongs to the field of the molecular biology, and relates to the traditional Chinese medicine and traditional Chinese medicinal material identification field. The kit comprises a solution A, a solution B, a reagent C and a user manual; and the solution A is composed of a solute triton-100 and a solvent water, the concentration of the solute is 1%, and the final concentration of the solute in the solution A 1%. The solution B is composed of 1M of a solute Tris-HCl and the solvent water, the concentration of the Tris-HCl in the solution B is 1M, the pH value of the solution B is 8.0, the concentration of the solute is 0.1M, and the final solution of the solute in the solution B is 0.1M. The reagent C is composed of NaOH and polyvinylpyrrolidone (PVP, M=40), the final concentration of NaOH in the solution A is 0.5M, and the final concentration of the PVP in the solution A is 1%. The kit is a medicinal material genome DNA extraction product developed for rapidly discriminating the medicinal material, and can be used for rapidly extracting plant and animal medicinal material genome DNA. The kit has the advantages of few operation steps, fast extraction speed, low price, no need of a large device, large-scale high-flux extraction and the like, and is very suitable for rapid detection and identification of the medicinal material DNA.
Description
technical field [0001] The invention relates to the field of identification of traditional Chinese medicines and Chinese medicinal materials, in particular to a rapid DNA extraction kit for medicinal materials. Belonging to the field of molecular biology technology Background technique [0002] For a long time, DNA extraction has been the most time-consuming and cumbersome step in DNA analysis technology experiments, which seriously restricts the use and promotion of molecular identification methods. In order to achieve the purpose of rapid DNA extraction, researchers have successively proposed ROSE method, magnetic bead adsorption method, Tris-EDTA extraction method, etc. for the extraction of plant DNA. Although these methods shorten the time of DNA extraction, they still cannot achieve the purpose of extracting DNA quickly and easily because they still use steps such as heating, high-speed centrifugation, and precipitation. Among them, the DNA alkaline lysis method is c...
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IPC IPC(8): C12N15/10
Inventor 黄璐琦袁媛蒋超李曼南铁贵
Owner INST OF CHINESE MATERIA MEDICA CHINA ACAD OF CHINESE MEDICAL SCI
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