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The detection method of Radix Paeoniae Alba

A detection method and technology of Radix Paeoniae Alba, applied in the direction of biochemical equipment and methods, microbiological determination/inspection, etc., can solve the problem that it is difficult to accurately distinguish authentic Radix Radix Radix Albae Accurate classification and good repeatability

Active Publication Date: 2018-12-28
GUANGZHOU XIANGXUE PHARMA CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The appearance, texture and texture of authentic white peony and non-authentic white peony are very similar, and it is difficult to accurately distinguish authentic white peony from non-authentic white peony by using traditional methods

Method used

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  • The detection method of Radix Paeoniae Alba
  • The detection method of Radix Paeoniae Alba
  • The detection method of Radix Paeoniae Alba

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] A kind of Radix Paeoniae Alba variety detection method, comprises the following steps:

[0046] 1. Establish an identification model.

[0047] 1. Take different varieties of Radix Paeoniae Alba as reference substances, and the excavated reference species are 4-year-old sexually propagated Radix Paeoniae Alba and 4-year-old vegetatively propagated Radix Paeoniae Alba.

[0048] Sexually propagated Radix Paeoniae Alba is a sample planted in Shibali planting base of Huqiao Company. It is a variety imported from Shandong. There are 5 trees in total, marked as base No. 1, base No. 2, base No. 3, base No. 4, and base 5 No.

[0049] The vegetative propagation of Paeoniae Alba is a sample from the planting site of Shibali retail households. There are 5 local varieties of Paeoniae Alba in Bozhou that are propagated by buds, and they are marked as No. 1, No. 2, No. 3, No. 4, and No. 5 respectively.

[0050] 2. Extracting the genomic DNA of Radix Paeoniae Alba of the above-mentio...

Embodiment 2

[0089] A method for detecting Radix Paeoniae Alba varieties is basically similar to the method in Example 1, except that:

[0090] In order to obtain amplification products, the components of the polymerase chain reaction system used are:

[0091] The polymerase chain reaction volume was 25 μL. The components of the reaction system are: 2.5 μL 10× buffer buffer (i.e. DNA gel loading buffer), 1.0 μL MgCl with a concentration of 25 mmol / L 2 , 0.3 μL of dNTPs with a concentration of 10 mmol / L, 0.5 μL of primers with a concentration of 10 μmol / L, 20 ng of the DNA template obtained above, 0.1 μL of DNA polymerase with a concentration of 5 U / μL, and make up the remaining volume with ultrapure water.

[0092] The amplified products obtained above were electrophoretically separated by agarose gel electrophoresis. Compared with Example 1, the electrophoretic bands were less and blurred, making it difficult to perform subsequent cluster analysis and calculation of similarity coefficien...

Embodiment 3

[0094] A method for detecting Radix Paeoniae Alba varieties is basically similar to the method in Example 1, except that:

[0095] In order to obtain amplification products, the components of the polymerase chain reaction system used are:

[0096]The polymerase chain reaction volume was 25 μL. The components of the reaction system are: 2.5 μL 10× buffer buffer (i.e. DNA gel loading buffer), 2.5 μL MgCl with a concentration of 25 mmol / L 2 , 0.5 μL of dNTPs with a concentration of 10 mmol / L, 1.25 μL of primers with a concentration of 10 μmol / L, 50 ng of the DNA template obtained above, 0.1 μL of DNA polymerase with a concentration of 5 U / μL, and make up the remaining volume with ultrapure water.

[0097] The amplified products obtained above were electrophoretically separated by agarose gel electrophoresis. Compared with Example 1, the electrophoretic bands were blurred, and it was difficult to perform subsequent clustering analysis and calculation of similarity coefficient.

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Abstract

The invention relates to a white peony root variety identifying method and belongs to the technical field of plant variety identification. The method includes: taking white peony roots of different varieties, respectively extracting the DNA of the white peony roots, using primers to perform polymerase chain reaction amplification on the extracted DNA to obtain amplification products, performing electrophoretic separation on the amplification products to obtain the DNA electrophoretic graphs of the white peony roots of different varieties, and building a white peony root variety identifying model according to the spectrogram information of the electrophoretic graphs; taking to-be-identified white peony roots, acquiring the DNA electrophoretic graphs of the white peony roots as before, substituting the spectrogram information of the electrophoretic graphs of the to-be-identified white peony roots into the white peony root variety identifying model for analysis so as to identify the varieties of the to-be-identified white peony roots. The method provides an effective and reliable white peony root variety identifying manner and has huge advantages as compared with traditional morphological identification, and a new measure for solving variety chaos and determining variety advantages and disadvantages in white peony root researches and application is provided.

Description

technical field [0001] The invention relates to a method for identifying plant varieties, in particular to a method for detecting varieties of Radix Paeoniae Alba. Background technique [0002] ISSR (inter-simple sequence repeat) is a molecular marker technology based on microsatellites developed by Zietkeiwitchz in 1994. The basic principle is to use the anchored microsatellite DNA as a primer, that is, add 2-4 random nucleotides to the 3' end or 5' end of the SSR sequence. In the PCR reaction, the anchor primer can cause a specific site Annealing, resulting in PCR amplification of a not too widely spaced inter-repeat DNA fragment complementary to the anchor primer. Multiple bands in the amplified inter SSR region were resolved by polyacrylamide gel electrophoresis, and the amplified bands were mostly dominant. [0003] According to modern molecular biology studies, the diversity of biological germplasm resources of Chinese herbal medicines (excluding mineral medicines) i...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6895
CPCC12Q1/6827C12Q1/6895C12Q2600/13C12Q2600/156
Inventor 胡珊王永辉连林生张健陈洪毅
Owner GUANGZHOU XIANGXUE PHARMA CO LTD