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Method for extracting DNA from deep process type traditional Chinese medicine or traditional Chinese medicinal materials

A processing technology of traditional Chinese medicinal materials, applied in DNA preparation, recombinant DNA technology, biochemical equipment and methods, etc., can solve the problems of short fragments, difficult molecular biology detection, difficult to obtain DNA samples, etc., to reduce losses. Effect

Active Publication Date: 2008-09-24
EAST CHINA UNIV OF SCI & TECH +1
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0012] It can be found that we may be able to obtain a small amount of DNA samples by simply using any of the commonly used DNA extraction methods that currently exist to extract nucleic acids from gelatinous traditional Chinese medicines, but it is very difficult to continue to apply them to subsequent molecular biology tests. In fact, if we directly use various domestic and foreign DNA extraction commercial kits currently on the market for DNA extraction of donkey-hide gelatin, it is also difficult for us to obtain DNA samples that can meet the requirements of subsequent testing.
Because it is inevitable, none of these methods can effectively address the characteristics of high protein content, low DNA content, and short fragments in colloidal traditional Chinese medicine.

Method used

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  • Method for extracting DNA from deep process type traditional Chinese medicine or traditional Chinese medicinal materials
  • Method for extracting DNA from deep process type traditional Chinese medicine or traditional Chinese medicinal materials
  • Method for extracting DNA from deep process type traditional Chinese medicine or traditional Chinese medicinal materials

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0104] Example 1 extracts the contained DNA in donkey-hide gelatin and carries out PCR detection to it

[0105] Commercially available donkey-hide gelatin (Shandong Dong'e donkey-hide gelatin Co., Ltd.) was used as the material, and the DNA was extracted using the following method.

[0106] 1) The solid colloid sample to be tested is hammered and pulverized into granules.

[0107] 2) Take 1g of pulverized glue sample, add 1ml of digestion buffer, seal in a 60ml centrifuge tube, and keep it in a water bath at 50-60°C for several minutes, stirring gently from time to time until all the granular substances are dissolved.

[0108] 3) After cooling to room temperature, 50 μl of 20 mg / ml proteinase K solution was added, mixed well, and kept at 56° C. for 1 hour, with slight shaking from time to time.

[0109] 4) Add 10ml PN buffer and mix well.

[0110] 5) Take 700 μl of the mixed solution into an adsorption column, centrifuge at 13,000 g at room temperature for 1 min, and discard...

Embodiment 2

[0122] Embodiment 2 extracts the DNA contained in yellow gelatin and carries out PCR detection to it

[0123] Using commercially available yellow gelatin (Shandong Dong'e Ejiao Co., Ltd.) as a raw material, DNA was extracted using the following method.

[0124] 1) The solid colloid sample to be tested is hammered and pulverized into granules.

[0125] 2) Take 1g of pulverized glue sample, add 1ml of digestion buffer, seal in a 60ml centrifuge tube, and keep it in a water bath at 50-60°C for several minutes, stirring gently from time to time until all the granular substances are dissolved.

[0126] 3) After cooling to room temperature, 50 μl of 20 mg / ml proteinase K solution was added, mixed well, and kept at 56° C. for 1 hour, with slight shaking from time to time.

[0127] 4) Add 10ml PN buffer and mix well.

[0128] 5) Take 700 μl of the mixed solution into an adsorption column, centrifuge at 13,000 g at room temperature for 1 min, and discard the collected solution.

[0...

Embodiment 3

[0140] Embodiment 3 extracts the DNA contained in the pig skin glue and carries out PCR detection to it

[0141] The following method was used to extract the DNA from pigskin glue made from pigskin by Shandong Dong'e Ejiao Co., Ltd. Research Institute.

[0142] 1) The solid colloid sample to be tested is hammered and pulverized into granules.

[0143] 2) Take 1g of pulverized glue sample, add 1ml of digestion buffer, seal in a 60ml centrifuge tube, and keep it in a water bath at 50-60°C for several minutes, stirring gently from time to time until all the granular substances are dissolved.

[0144] 3) After cooling to room temperature, 50 μl of 20 mg / ml proteinase K solution was added, mixed well, and kept at 56° C. for 1 hour, with slight shaking from time to time.

[0145] 4) Add 10ml PN buffer solution, mix evenly, add 2ml n-hexane, slightly invert and mix well, centrifuge at 3000g, 4°C for 5min, discard the supernatant, and transfer the lower solution to a clean 50ml centr...

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Abstract

The invention provides a method for extracting DNA from deeply processed Chinese traditional medicines or Chinese traditional medicinal materials. The method comprises the steps of: a) pretreating the deeply processed Chinese traditional medicines or Chinese traditional medicinal materials with digestive juice, digesting the Chinese traditional medicines or Chinese traditional medicinal materials with 0.5 percent of SDS and a proteolytic enzyme K so that a lot of collagen in the deeply processed Chinese traditional medicines or Chinese traditional medicinal materials is degraded, and little DNA is completely released and dissolved in the digestive juice to get pretreatment solution; b) extracting DNA in the pretreatment solution with the extraction method of short fragment DNA. The method is quick, simple, convenient, reliable and efficient, which is in particular suitable to glue-type Chinese traditional medicines containing little nucleic acid and very short fragments resulting from deeply processing; the acquired DNA can be used for subsequent general molecular biology operations. The method provided by the invention provides a base for distinguishing glue-type Chinese traditional medicines with the DNA biologic identification technology.

Description

technical field [0001] The present invention relates to the technical field of traditional Chinese medicine, more specifically, to the technical field of DNA extraction from traditional Chinese medicine, in particular to a method for extracting DNA from deep-processed traditional Chinese medicine or Chinese medicinal materials, which is very suitable for deep-processed products, especially because Deep processing results in Chinese herbal medicines or Chinese herbal medicines with very little DNA content and extremely short fragments, especially gelatinous Chinese medicines or Chinese herbal medicines refined from animal skins and bones. Background technique [0002] my country's traditional Chinese medicine is extensive and profound, and there are many kinds of substances that can be used as Chinese medicine. One of them is glue-like Chinese medicine made from animal skin and bone, including donkey-hide gelatin, yellow gelatin, tortoise shell glue, and deer horn glue. These...

Claims

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Application Information

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IPC IPC(8): C12N15/10C12Q1/68
Inventor 周祥山张元兴吕品杨继忠秦玉峰尤金花田守生
Owner EAST CHINA UNIV OF SCI & TECH
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