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Specific primer pair and method thereof for identifying acorus calamus L. and acorus tatarinowii schott

A technique for Tibetan calamus and Shi calamus, which is applied in the field of identifying specific primer pairs of Tibetan calamus and Shi calamus respectively, can solve the problems of carcinogenicity, teratogenicity, confusion of names, etc., and achieve the effect of ensuring the quality of medicinal materials

Active Publication Date: 2015-12-23
HUNAN INST FOR FOOD & DRUG CONTROL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the toxicity of α-asarone and β-asarone, the active ingredients of Tibetan calamus, to humans, the U.S. FDA (Food and Drug Administration) has banned the consumption of Tibetan calamus and its extracts.
Historically, the use of Tibetan calamus and stone calamus has not been clearly separated, and due to the wide range of regions, there are too many common names, which leads to confusion in names

Method used

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  • Specific primer pair and method thereof for identifying acorus calamus L. and acorus tatarinowii schott
  • Specific primer pair and method thereof for identifying acorus calamus L. and acorus tatarinowii schott
  • Specific primer pair and method thereof for identifying acorus calamus L. and acorus tatarinowii schott

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] The method for identifying Tibetan calamus and Shichangpu includes the following steps:

[0044] 1. Extraction of medicinal DNA: proceed as usual, adjust the DNA concentration of the test sample to 0.2~0.5μg / μL with deionized water;

[0045] 2. Polymerase chain reaction:

[0046] (1) The polymerase chain reaction is based on 25μL, and the dosages of various items are:

[0047]

[0048]

[0049] (2) PCR reaction conditions: the reaction was performed on a PCR machine, and the reaction conditions were: 94°C pre-denaturation for 5 minutes, 94°C denaturation for 30s, 62°C annealing for 30s, 72°C extension for 45s, and after 35 cycles, 72°C extension for 10 minutes.

[0050] (3) Electrophoresis detection of PCR products: Take 5μL of the above reaction solution, mix it with 1.0μL loading buffer, and use 2% agarose gel (containing 0.5μg / μLGelRed) to detect the amplification results by electrophoresis. All DNA samples can be Amplify a DNA band of about 317bp;

[0051] 3. Restriction enzy...

Embodiment 2

[0059] In order to ensure the reliability of the molecular identification results, this experiment collected a variety of fake Tibetan calamus and Shichangpu and 50 common Chinese medicinal materials circulating in the market to verify the applicability of the experimental method to ensure that the method is accurate and reliable .

[0060] 1 Materials (see Table 1)

[0061] Table 1 Source of all samples of the present invention

[0062]

[0063]

[0064] 2DNA extraction

[0065] Take 0.1g of this product, wash with 1ml of 75% ethanol and 1ml of sterilized ultrapure water in turn, absorb the surface moisture, and grind it into a very fine powder in a mortar. Take 25mg, put it in a 1.5ml centrifuge tube, extract DNA with the new broad-spectrum plant genomic DNA rapid extraction kit, add 0.7mL 65℃ preheating buffer GP1, add mercaptoethanol to make the concentration 0.1%, mix gently, Place the centrifuge tube in a 65°C water bath for 20 minutes, shaking occasionally. Add 0.7 mL of chl...

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Abstract

The invention discloses a specific primer pair and a method thereof for identifying acorus calamus L. and acorus tatarinowii schott. The primer pair includes a primer P1 and a primer P2, wherein the sequence of the primer P1 is as shown in SEQ ID NO.1 and the sequence of the primer P2 is as shown in SEQ ID NO.2. The method can be used for respectively identifying acorus calamus L. and acorus tatarinowii schott by carrying out simple DNA extraction, PCR specific amplification, enzyme digestion and electrophoresis detection on samples; the method has the advantages of simple operation, strong specificity, good repeatability and the like; and the method is applicable to rapid identification of acorus calamus L. and acorus tatarinowii schott medicinal materials.

Description

Technical field [0001] The invention relates to the technical field of quality identification of Chinese medicinal material germplasm, in particular to a specific primer pair for identifying Tibetan calamus and Shichangpu and a method thereof. Background technique [0002] Tibetan calamus and Shichangpu are the species included in the 2010 edition of the Chinese Pharmacopoeia. Tibetan calamus is the dried rhizome of Acorus calamus Acoruscalamus L., and Acorus calamus is the dried rhizome of Acorustatarinowii Schott. Traditional Tibetan medicinal materials. Tibetan calamus and Shichangpu medicinal materials are widely used in our country, and they are important species with the same medicine and food. They were first published in "Shen Nong's Materia Medica" and have been used today. In Tibetan areas, the use and history of Tibetan calamus has great value. It is classified as a first-class endangered Tibetan medicinal material, and the medicinal resources are lacking. It has the...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/686C12Q1/6895C12Q2521/301C12Q2563/185
Inventor 聂平肖炳燚罗晖明李文莉刘丽丁野舒毕琼孙辉
Owner HUNAN INST FOR FOOD & DRUG CONTROL
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