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Method for extracting agilawood high-quality DNA

A high-quality, agarwood technology, applied in the fields of molecular biology and traditional Chinese medicine identification, can solve the problems of difficulty in extracting high-purity DNA, poor extraction effect, and high cost of agarwood, to improve DNA extraction efficiency, improve quantity and quality, and avoid problems. Extraction of unstable effects

Active Publication Date: 2018-10-09
INST OF MEDICINAL PLANT DEV CHINESE ACADEMY OF MEDICAL SCI HAINAN BRANCH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The extraction of its high-purity DNA is difficult, and the extraction effect of conventional methods is poor, and it is difficult to meet the requirements in the fields of germplasm resources, aroma mechanism, species identification, genetic evolution, phylogeny, and variety selection. , it is often necessary to obtain high-quality DNA from agarwood, so the extraction of high-quality DNA is an essential step
In addition, the cost of agarwood is too high, and it is expensive to re-purchase experimental agarwood samples

Method used

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  • Method for extracting agilawood high-quality DNA
  • Method for extracting agilawood high-quality DNA
  • Method for extracting agilawood high-quality DNA

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] A method for extracting high-quality DNA from agarwood, the steps of which are:

[0044] (1) Take 200 mg of dry Acacia wood, wipe the surface with 75% alcohol to remove surface dirt, dry it, chop it up, put it in a 5 mL centrifuge tube, add liquid nitrogen to fully seal and grind for 5 min.

[0045] (2) Quickly add 1000 μL of preheated DNA extraction solution and shake well; incubate in a 60°C water bath for 8 hours, shaking the centrifuge tube several times during this period; add an equal volume of DNA separation solution, vortex for 30 seconds, and centrifuge at 9000 rpm for 20 minutes at room temperature.

[0046] (3) Transfer the supernatant obtained in step (2) to a new 5mL centrifuge tube to ensure that no residue is sucked in; add 2 times the volume of DNA precipitation solution, mix well by suction, and stand at -20°C for 18h.

[0047] (4) Take out the centrifuge tube from step (3) and cool it at room temperature, add an equal volume of DNA detergent, put it in a...

Embodiment 2

[0069] A method for extracting high-quality DNA from agarwood, the steps of which are:

[0070] (1) Take 200 mg of dried Acacia wood, wipe the surface with bleach and remove the surface dirt, dry it, chop it and place it in a 5 mL centrifuge tube, add liquid nitrogen to fully seal and grind for 5 min.

[0071] (2) Quickly add 2000 μL of preheated DNA extraction solution and shake well; incubate in a 50°C water bath for 3 hours, shaking the centrifuge tube several times during this period; add an equal volume of DNA separation solution, vortex for 30 seconds, and centrifuge at 4500 rpm for 1 minute at room temperature.

[0072] (3) Transfer the supernatant obtained in step (2) to a new 5mL centrifuge tube to ensure that no residue is sucked in; add 2 times the volume of DNA precipitation solution, mix well by suction, and stand at -20°C for 12h.

[0073] (4) Take out the centrifuge tube from step (3) and cool it at room temperature, add an equal volume of DNA detergent, put it ...

Embodiment 3

[0088] A method for extracting high-quality DNA from agarwood, the steps of which are:

[0089] (1) Take 100mg of dry Akiras wood, wipe the surface with 75% alcohol to remove surface dirt, dry and chop, put in a 5mL centrifuge tube, add liquid nitrogen to fully seal and grind for 5min.

[0090] (2) Quickly add 500 μL of preheated DNA extraction solution and shake well. Warm in a water bath at 65°C for 8 hours, during which the centrifuge tubes were shaken several times. Add an equal volume of DNA separation solution, vortex for 30s, and centrifuge at 12000rpm for 1min at room temperature.

[0091] (3) Transfer the supernatant obtained in step (2) to a new 5mL centrifuge tube, making sure not to inhale the residue. Add 2 times the volume of DNA precipitation solution, mix with suction, and let stand at -20°C for 12h.

[0092] (4) Take out the centrifuge tube from step (3) and cool it at room temperature, add an equal volume of DNA detergent, put it in an ice bath for 10 minu...

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Abstract

The invention discloses a method for extracting agilawood high-quality DNA. The method includes steps: agilawood sample disinfection and impurity removal, liquid nitrogen grinding, warm bath, centrifuging, alcohol precipitation, washing dissolution and directional PCR amplification. High-quality DNA can be extracted from agilawood containing secondary metabolism substances like resin, especially from deeply-processed products (or residue) with high resin content and long formation (storage) time. Quality and quantity of agilawood DNA are improved, DNA can be extracted from the deeply-processedresidue, the method can be directly applied in studying molecular biology and traditional Chinese medicine identification science, and a new method is provided for agilawood species identification.

Description

technical field [0001] The invention belongs to the fields of molecular biology and identification of traditional Chinese medicines, and relates to a method for extracting DNA from agarwood, in particular to a method for extracting DNA from agarwood with high resin content, long formation (storage) time, and deep-processing product (or residue). Background technique [0002] Agarwood is a wood containing resin in the genus Agarwood or Pseudo-Agarwood of the family Daphneaceae, and is a precious traditional Chinese medicine. As a medicine, agarwood was first recorded in "Famous Doctors" written by Tao Hongjing in the Liang Dynasty. disease. In the process of circulation, information such as the species of agarwood and the way of agarwood is missing, and then processed, so it needs to be identified to judge its quality. The traditional method of "seeing, smelling, and burning" has subjective consciousness, and the detection method of agarwood based on the "Chinese Pharmacopo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10
CPCC12N15/1003
Inventor 魏建和冯剑刘洋洋杨云康勇
Owner INST OF MEDICINAL PLANT DEV CHINESE ACADEMY OF MEDICAL SCI HAINAN BRANCH
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