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Method and kit for identifying genuine medicinal materials herba cynomorii different in growing area

An identification method and kit technology, applied in biochemical equipment and methods, microbial determination/inspection, DNA/RNA fragments, etc., can solve the problems of differentiation and identification difficulties, and achieve rapid identification, wide adaptability, The effect of high amplification efficiency

Active Publication Date: 2017-06-13
INST OF MEDICINAL PLANT DEV CHINESE ACADEMY OF MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Cynomorium cynomorium from different origins are similar in appearance and traits, and it is difficult to distinguish them by traditional morphological identification, especially for the identification of medicinal powder, decoction pieces, and Chinese patent medicines.

Method used

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  • Method and kit for identifying genuine medicinal materials herba cynomorii different in growing area
  • Method and kit for identifying genuine medicinal materials herba cynomorii different in growing area
  • Method and kit for identifying genuine medicinal materials herba cynomorii different in growing area

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] The identification method of Cynomorium cynomorium raw material in different origins of embodiment 1

[0026] 1) Collect 46 samples of Cynomorium cynomorae from different origins (see Table 1), take 25 mg of medicinal materials and grind them on MM400 ball mill (Retsch, Germany) respectively, and use the plant genome DNA reagent of Tiangen Biochemical Technology (Beijing) Co., Ltd. cassette to extract total DNA.

[0027] Table 1 Cynomorium samples from different origins

[0028]

[0029] 2) PCR amplification

[0030] The primer sequence is SY1F 5'-AAGGAAGCAGCAGCACATTGAGT-3',

[0031] SY1R 5'-AACCTGCGGAAGGATCATTGTTG-3',

[0032] Synthesized by Shanghai Meiji Biomedical Technology Co., Ltd. (Beijing).

[0033] The PCR amplification program is:

[0034]

[0035]

[0036] Above-mentioned PCR reaction system is:

[0037] PCR Buffer (10×) 2.5u L, Mg 2+ 2uL (25mmol / L), 2uL (2.5mmol / L) of dNTPs mixture, 1uL (2.5umol / L) of each primer, 2uL (about 30ng) of templat...

Embodiment 2

[0046] The identification method of embodiment 2 Cynomorium decoction pieces

[0047] Basically the same as Example 1, the difference is that Cynomorium cynomorium decoction pieces from different origins are used as samples, DNA is extracted and identified, and the results can also specifically detect the above 3 SNP sites, all Tacheng Cynomorium cypridis SEQ ID The 188th Tacheng Cynomorium from the 5' end of NO.1 sequence is T, and the 304th Tacheng Cynomorium is A, and the 305th Tacheng Cynomorium is T; the 305th Tacheng Cynomorium is C, Cynomorium from other places of origin is T. Sequencing results with figure 1 same.

Embodiment 3

[0048] The identification of embodiment 3 Cynomorium powder

[0049] Basically the same as Example 1, the difference is that Cynomorium Cymbidium powder from different origins is used as samples, DNA is extracted and identified, and the results can also specifically detect the above 3 SNP sites, all Tacheng Cynomorium Cynomorium SEQ ID The 188th Tacheng Cynomorium from the 5' end of NO.1 sequence is T, and the 304th Tacheng Cynomorium is A, and the 305th Tacheng Cynomorium is A; the 305th Tacheng Cynomorium is C, Cynomorium from other places of origin is T. Sequencing results with figure 1 same.

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Abstract

The invention relates to a method and kit for identifying genuine medicinal materials herba cynomorii different in growing area. The method comprises the steps that firstly, genome DNA of a to-be-detected sample serves as a template, and the segment containing the sequence shown in SEQ ID NO.1 is amplified; secondly, an amplified product is sequenced, after splicing, a primer area is deleted so as to obtain a complete amplified segment containing the sequence shown in SEQ ID NO.1, and basic groups at the 188th position and the 304th or 305th position starting from the 5' terminal in the sequence shown in SEQ ID NO.1 are detected; if the basic group at the 188th position starting from the 5' terminal in the sequence shown in SEQ ID NO.1 is T, or the basic group at the 304th position is A, or the basic group at the 305th position is C, the herba cynomorii is identified as Tacheng herba cynomorii, and if the basic group at the 188th position starting from the 5' terminal in the sequence shown in SEQ ID NO.1 is C, or the basic group at the 304th position is T, or the basic group at the 305th position is T, the herba cynomorii is identified as herba cynomorii from other growing areas. The method is wider in adaptability, high in primer specificity, high in amplification efficiency and capable of detecting any samples of herba cynomorii from which DNA can be extracted.

Description

technical field [0001] The invention belongs to the technical field of identification of source varieties of traditional Chinese medicinal materials, and in particular relates to an identification method for authentic medicinal materials Cynomorium cynomorae from different origins by using specific SNP markers. Background technique [0002] Cynomorium songaricum Rupr. is the dry fleshy stem of Cynomorium songaricum Rupr., also known as elixir, Ulan high waist, ground hair ball, sheep shaobula and so on. The fleshy stems of the inflorescences are removed for medicinal purposes. The fleshy stem is rich in tannins, which can be extracted from tannins, and contains starch, and can be used for wine making, feed and food substitutes. It has the functions of invigorating the kidney, benefiting essence, moistening dryness, etc. It is mainly used to treat impotence, nocturnal emission, soreness of the waist and knees, intestinal dryness and constipation, irregular menstruation, infe...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6895C12Q2600/156
Inventor 黄林芳李琳刘德旺陈士林
Owner INST OF MEDICINAL PLANT DEV CHINESE ACADEMY OF MEDICAL SCI
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