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Primer pair used for identification of traditional Chinese medicine bulbus fritillariae thunbergii, and applications thereof

A technique of primer pair and fritillary fritillary, applied in the field of molecular biology, can solve the problems of large differences in chemical components of fritillary plants, difficulties in identification by physical and chemical methods, and difficulty in identification, achieving good specificity, high sensitivity, and repeatable results good sex effect

Inactive Publication Date: 2017-09-15
SUZHOU LI LIANGJI HEALTH IND LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Variety confusion is a problem that is difficult to overcome in the practical application of fritillaria at present, and the traditional identification methods are subject to certain degree, especially for some fresh fritillary medicinal materials before flowering, dried or broken medicinal materials, when identifying It is more difficult, not only that, the chemical composition of Fritillaria species is quite different, and it is also difficult to identify by physical and chemical methods

Method used

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  • Primer pair used for identification of traditional Chinese medicine bulbus fritillariae thunbergii, and applications thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] A primer pair for identifying Fritillaria fritillata, said primer pair and its sequence are: P1, SEQ ID NO.1-2; P2, SEQ ID NO.3-4; P3, SEQ ID NO.5-6 .

[0026] A specific GC hairpin structure is added to the 5' end of the upstream primers of the primer pair P1, P2 and P3, the length of the hairpin structure is 40bp, and the sequence is SEQ ID NO.7.

[0027] Application of the primer pair in preparing a kit for identifying traditional Chinese medicine Zhe Fritillaria.

[0028] The components of the kit include: primer pairs, dNTP, DNA polymerase, LC Green, and reaction buffer.

[0029] The steps for identifying the traditional Chinese medicine Fritillaria fritillaria with the kit are:

[0030] (1) extract the genomic DNA of Fritillaria fritillata sample;

[0031] (2) using the genomic DNA extracted in step (1) as a template and using P1 as a specific primer to perform the first round of PCR amplification;

[0032] (3) Get the amplified product of step (2), dilute it ...

Embodiment 2

[0038] A primer pair for identifying Fritillaria fritillata, said primer pair and its sequence are: P1, SEQ ID NO.1-2; P2, SEQ ID NO.3-4; P3, SEQ ID NO.5-6 .

[0039] A specific GC hairpin structure is added to the 5' end of the upstream primers of the primer pair P1, P2 and P3, the length of the hairpin structure is 40bp, and the sequence is SEQ ID NO.7.

[0040] Application of the primer pair in preparing a kit for identifying traditional Chinese medicine Zhe Fritillaria.

[0041] The components of the kit include: primer pairs, dNTP, DNA polymerase, LC Green, and reaction buffer.

[0042] The steps for identifying the traditional Chinese medicine Fritillaria fritillaria with the kit are:

[0043] (1) extract the genomic DNA of Fritillaria fritillata sample;

[0044] (2) using the genomic DNA extracted in step (1) as a template and using P1 as a specific primer to perform the first round of PCR amplification;

[0045] (3) Get the amplified product of step (2), dilute it ...

Embodiment 3

[0051] A primer pair for identifying Fritillaria fritillata, said primer pair and its sequence are: P1, SEQ ID NO.1-2; P2, SEQ ID NO.3-4; P3, SEQ ID NO.5-6 .

[0052] A specific GC hairpin structure is added to the 5' end of the upstream primers of the primer pair P1, P2 and P3, the length of the hairpin structure is 40bp, and the sequence is SEQ ID NO.7.

[0053] Application of the primer pair in preparing a kit for identifying traditional Chinese medicine Zhe Fritillaria.

[0054] The components of the kit include: primer pairs, dNTP, DNA polymerase, LC Green, and reaction buffer.

[0055] The steps for identifying the traditional Chinese medicine Fritillaria fritillaria with the kit are:

[0056] (1) extract the genomic DNA of Fritillaria fritillata sample;

[0057] (2) using the genomic DNA extracted in step (1) as a template and using P1 as a specific primer to perform the first round of PCR amplification;

[0058] (3) Get the amplified product of step (2), dilute it ...

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Abstract

The invention discloses a primer pair used for identification of traditional Chinese medicine bulbus fritillariae thunbergii, and applications thereof. The sequences of the primer pair are represented by P1, SEQ ID NO.1-2; P2, SEQ ID NO.3-4; and P3, SEQ ID NO.5-6. The primer pair possesses following advantages: 1, the primer pair can be used for identification of single sample directly, an application method is simple in operation, result is accurate, and application range is wide; and 2, the primer pair is high in specificity, result repeatability, and sensitivity, and is suitable to be used for producing detection kits.

Description

technical field [0001] The invention belongs to the field of molecular biology and relates to a method for identifying traditional Chinese medicine by means of molecular biology, in particular to a pair of primers for identifying fritillary fritillaria and its application. Background technique [0002] Fritillaria fritillaria is one of the authentic medicinal materials "Eight Flavors of Zhejiang Province". It has the effect of nourishing the lungs and relieving cough, and has a long history of medication. There are many varieties of Fritillaria. In addition to Zhejiang Fritillaria, the main varieties of Fritillaria include Sichuan Fritillaria, Hubei Fritillaria, Flat Fritillary and Yi Fritillaria. In addition, there are many counterfeit Fritillaria in the circulation field. The medicinal effects of different varieties of Fritillaria are significantly different, and the price varies greatly, so it is necessary to establish an accurate and efficient identification method. At ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6895
Inventor 李建华
Owner SUZHOU LI LIANGJI HEALTH IND LTD
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