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A dna hybridization method for identifying American ginseng

A hybridization method, the technology of American ginseng, which is applied in the field of DNA hybridization for identifying American ginseng, can solve the problems of taking a lot of time and cost

Active Publication Date: 2022-05-27
珠海澳加动力生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the current DNA barcoding technology used to identify American ginseng will include two steps of PCR product formation and sequence identification, and both involve gel electrophoretic separation. The process involves electric field action and special staining, which takes a lot of time and operation Technical cost, the demand for rapid identification of American ginseng is still unsatisfactory, so it is necessary to provide a technology for rapid identification of American ginseng

Method used

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  • A dna hybridization method for identifying American ginseng
  • A dna hybridization method for identifying American ginseng
  • A dna hybridization method for identifying American ginseng

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] 1. Various solution configurations

[0052] Configuration of Probe Solutions

[0053] Take the dissolving solution and the probe, and configure the probe solution. The components of the configured probe solution are as follows:

[0054] (1) 25 μM amine-labeled oligonucleotide single-stranded

[0055] (2) 1.5M NaCl

[0056] (3) 0.15M sodium bicarbonate

[0057] (4) 0.15% sodium dodecyl sulfonate (SDS)

[0058]Configuration of the DNA solution of the sample to be tested

[0059] Take the lysate and the DNA of the sample to be tested, and configure the DNA solution of the sample to be tested. The components of the prepared DNA solution of the sample to be tested are as follows:

[0060] (1) 25nM biotin-labeled oligonucleotide single-stranded

[0061] (2) 0.15M NaCl

[0062] (3) 0.015M sodium citrate (pH 7.0)

[0063] (4) 0.15% sodium dodecyl sulfonate (SDS)

[0064] Configuration of Gold Nanoparticle (AuNP) Solutions

[0065] Before the experiment, add the AuNP s...

Embodiment 2

[0076] Example 2 Screening of probe sequences

[0077] Two probes were designed, denoted as N1Q and N2Q, respectively, and the sequences of the two probes are:

[0078] N1Q: CTAAAAAAAAAGTATTTCTCATCTAAATTTTGAA (SEQ ID NO: 1)

[0079] N2Q: GAATTTGAAAGTGTTTTAAATTGATTTTCAA (SEQ ID NO: 2)

[0080] Two DNA samples were prepared, which were derived from Chinese ginseng and American ginseng, respectively.

[0081] The experiment was carried out according to the operation of Example 1, wherein step (4) in Example 1 used 1 times PBS buffer for washing the microfluidic chip, and the experimental results were as follows image 3 shown.

[0082] from image 3 It can be seen from the above that in the reaction area where the probe N2Q is fixed, there is no significant difference in the fluorescence reaction of Chinese ginseng and American ginseng, and the probe N2Q cannot distinguish Chinese ginseng and American ginseng well; while in the reaction area where the probe N1Q is fixed, Amer...

Embodiment 3

[0083] Example 3 Screening of gold nanoparticle solutions

[0084] Experiment A

[0085] Two DNA samples were prepared, which were derived from Chinese ginseng and American ginseng respectively.

[0086] The experiment was carried out according to the operation of Example 1, wherein the washing solution in step (4) washing the microfluidic chip in Example 1 was: (1) Group 1 used 1 times PBS buffer; (2) Group 2 used gold The solution with 5 nM nanoparticles concentration; (3) the solution with 10 nM gold nanoparticles concentration used in group 3; (4) the solution with 15 nM gold nanoparticles concentration used in group 4; (5) the solution used in group 5 A solution with a gold nanoparticle concentration of 20 nM; the experimental results are as follows Figure 4 shown.

[0087] 1, 3, 5, 7 and 9 in the longitudinal direction are ginseng reaction areas, and 2, 4, 6, 8 and 10 are American ginseng reaction areas.

[0088] Groups 1-5, the corrected fluorescence values ​​of ea...

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Abstract

The probe used in the DNA hybridization method for identifying American ginseng provided by the present invention is based on the single nucleotide polymorphism (Single Nucleotide Polymorphism, SNP) site of the dammarenediol synthase (dammarenediol synthase) gene in the Panax genus genome design. The inventor found that the dammarene diol synthase genes of Chinese ginseng, American ginseng, and Panax notoginseng have high similarity in experiments. In order to more accurately identify American ginseng, the inventor designed a variety of probe sequences, and finally screened out the dammarene diol synthase that can be accurately identified. The probe sequence of American ginseng.

Description

technical field [0001] The present invention relates to a DNA hybridization method for identifying American ginseng. Background technique [0002] Chinese herbal medicine ginseng belongs to Plantae (plant kingdom), Apiales (Umbelliferae), Araliaceae (Araliaceae), Aralioideae (Ginseng subfamily), Panax (Ginseng genus) in taxonomy. There are three main species classifications of ginseng: Panax Ginseng (Asian ginseng / Chinese ginseng), Panax notoginseng (Panax notoginseng) and Panax Quinquefolius (American ginseng). [0003] Although Chinese ginseng, Panax notoginseng and American ginseng originate from the same genus, the uses and properties of the three herbs are different. For example, Chinese ginseng does not affect the effect of warfarin drugs, but American ginseng can significantly affect the anticoagulant activity of warfarin. [0004] Because Chinese ginseng, Panax notoginseng and American ginseng have different functions and values, the technology and means of rapid i...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6895C12Q1/6837
CPCC12Q1/6837C12Q1/6895C12Q2563/131
Inventor 李志恒奥伯克·克里斯托弗
Owner 珠海澳加动力生物科技有限公司
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