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Medicinal liquorice root consanguinity identification method based on ITS2 sequence

A technology of genetic relationship and identification method, which is applied in the field of genetic relationship identification of medicinal licorice based on the ITS2 sequence, can solve problems such as confusion in medicinal use, difficulty in distinguishing licorice species, and affecting the effectiveness and safety of clinical medication.

Inactive Publication Date: 2017-10-03
GUIYANG COLLEGE OF TRADITIONAL CHINESE MEDICINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Therefore, in the prior art, the licorice species in the genus Glycyrrhiza are not easy to distinguish, which makes the medicinal situation of the licorice market chaotic, seriously affecting the effectiveness and safety of clinical medication.

Method used

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  • Medicinal liquorice root consanguinity identification method based on ITS2 sequence
  • Medicinal liquorice root consanguinity identification method based on ITS2 sequence
  • Medicinal liquorice root consanguinity identification method based on ITS2 sequence

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0067] Genetic relationship identification method of medicinal licorice based on ITS2 sequence; including DNA extraction, PCR amplification, sequencing and determination of licorice genetic relationship;

[0068] The DNA extraction is to take the licorice sample tissue, wipe the surface with alcohol and dry it, and extract the total DNA with the plant genome DNA extraction kit. The water bath time is 60min during the extraction process, and it is diluted to 8-12ng·μL -1 , -22--18 ℃ storage;

[0069] The PCR amplification is to perform an amplification reaction with a forward primer and a reverse primer to obtain an amplification product; the sequencing is to perform bidirectional sequencing after the amplification product is purified;

[0070] The forward primer; is ATG CGA TAC TTG GTG TGA AT; the concentration is 2.5 μmol L -1 ;

[0071] The reverse primer; is GAC GCT TCT CCA GAC TAC AAT; the concentration is 2.5 μmol L -1 ;

[0072] The amplification reaction described: ...

Embodiment 2

[0077] Genetic relationship identification method of medicinal licorice based on ITS2 sequence; including DNA extraction, PCR amplification, sequencing and determination of licorice genetic relationship;

[0078] The DNA extraction is to take the licorice sample tissue, wipe the surface with alcohol and dry it, and extract the total DNA with a plant genome DNA extraction kit. During the extraction process, the water bath time is 70min, and it is diluted to 12ng·μL -1 , -22--18 ℃ storage;

[0079] The PCR amplification is to perform an amplification reaction with a forward primer and a reverse primer to obtain an amplification product; the sequencing is to perform bidirectional sequencing on the amplification product after purification;

[0080] The forward primer; is ATG CGA TAC TTG GTG TGA AT; the concentration is 3.0 μmol L -1 ;

[0081] The reverse primer; is GAC GCT TCT CCA GAC TAC AAT; the concentration is 3.0 μmol L -1 ;

[0082] The amplification reaction described:...

Embodiment 3

[0088] Genetic relationship identification method of medicinal licorice based on ITS2 sequence; including DNA extraction, PCR amplification, sequencing and determination of licorice genetic relationship;

[0089] The DNA extraction is to take the licorice sample tissue, wipe the surface with alcohol and dry it, and extract the total DNA with a plant genome DNA extraction kit. During the extraction process, the water bath time is 50min, and it is diluted to 8ng·μL -1 , -22--18 ℃ storage;

[0090] The PCR amplification is to perform an amplification reaction with a forward primer and a reverse primer to obtain an amplification product; the sequencing is to perform bidirectional sequencing on the amplification product after purification;

[0091] The forward primer; is ATG CGA TAC TTG GTG TGA AT; the concentration is 2.0 μmol L -1 ;

[0092] The reverse primer; is GAC GCT TCT CCA GAC TAC AAT; the concentration is 2.0 μmol L -1 ;

[0093] The amplification reaction described: ...

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Abstract

The invention discloses a medicinal liquorice root consanguinity identification method based on an ITS2 sequence. The method includes the identification on the consanguinity of medicinal liquorice roots and sibling species samples. The glycyrrhiza liquorice root species can be identified, so that various liquorice root species can be easily distinguished; the liquorice root medicinal condition disorder condition si avoided; the support is provided for ensuring the effectiveness and safety of liquorice root clinic medicine use.

Description

technical field [0001] The invention relates to a method for identifying the genetic relationship of medicinal licorice, in particular to a method for identifying the genetic relationship of medicinal licorice based on an ITS2 sequence (an identification method for medicinal licorice based on the ITS2 sequence). Background technique [0002] Glycyrrhiza Glycyrrhiza Linn. has about 20 species worldwide. There are 8 species in my country, namely Glycyrrhiza glabra (G.uralensis), Glycyrrhiza glabra (G.glabra), also known as Glycyrrhiza glabra, G.inflata, Glycyrrhiza glabra (G.eglandulosa X.Y.Li.) , G. aspera Pall., and G. pallidiflora Maxim., G. yunnanensis Cheng f.et L.K.Dai ex P.C.Li, and G. squamulosa Franch. The 2015 edition of "The Pharmacopoeia of the People's Republic of China" clearly pointed out that the medicinal licorice in my country is the dried root and rhizome of licorice, glabra and bloated licorice, which have the functions of nourishing the spleen and replenis...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/6895C12Q2600/156
Inventor 刘育辰郝杰刘刚李悦袁思文金文渊
Owner GUIYANG COLLEGE OF TRADITIONAL CHINESE MEDICINE
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