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Periplaneta Americana L. specific COI primer, kit containing same and application of Periplaneta Americana L. specific COI primer

The technology of a Periplaneta americana and a kit, which is applied in the field of molecular biology, can solve problems such as identification errors, identification difficulties, and non-specificity of tissue characteristics, and achieve the effects of improving accuracy, strong specificity, and simple operation

Active Publication Date: 2017-09-19
SICHUAN GOODDOCTOR PANXI PHARMA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, since most insect medicinal materials have similar external shapes and non-specific tissue characteristics, and it is difficult to see the shape and color of the species after rough processing in the place of origin and processing of decoction pieces, which brings many difficulties to accurate medicinal material identification, so the traditional morphology There are problems of recognition difficulty and recognition error in the method of scientific feature recognition

Method used

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  • Periplaneta Americana L. specific COI primer, kit containing same and application of Periplaneta Americana L. specific COI primer
  • Periplaneta Americana L. specific COI primer, kit containing same and application of Periplaneta Americana L. specific COI primer

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Embodiment 1 Periplaneta americana-specific primers JmF1 and JmR1 are to the amplification effect of Periplaneta americana

[0026] 1. Pretreatment of the test product

[0027] Before the genome preparation of the test sample, the surface of the sample was first wiped with 75% ethanol to eliminate exogenous contamination. After the ethanol volatilized, the abdominal muscle tissue was selected and fully ground as the genome extraction sample.

[0028] 2. Preparation of the Periplaneta americana genome

[0029] Using a DNA extraction kit (TIANamp Genomic DNA Kit (Beijing Tiangen Biochemical Technology Co., Ltd.)) to extract the template DNA of the American cockroach, use a micro-spectrophotometer to detect the concentration of the extracted template DNA of the American cockroach, and use sterilized deionized water Dilute the DNA concentration of the sample to 0.1ug / ul-0.2ug / ul.

[0030] 3. Synthesis of Periplaneta americana-specific COI primer sequences

[0031] The se...

Embodiment 2

[0040] Example 2 Determination of the minimum detection amount of Periplaneta americana by primers JmF1 and JmR1.

[0041] Genomic DNA of Periplaneta americana was extracted according to Example 1, and amplified according to the system reaction system and PCR reaction conditions described in Example 1. Then the original template solution (DNA solution concentration is 50ng / ul) is carried out descending gradient dilution with 2 times, the solution after getting 2ul dilution is used as the template of PCR amplification, directly adds in the PCR reaction system, and the description of reaction system is the same as in Example 1 . Dilute until no bands are detected.

[0042] Utilize primer JmF1 and JmR1 (SEQ ID No.1 and SEQ ID No.2) to make the mensuration of minimum detection amount, carry out PCR amplification with the American cockroach genomic DNA of different dilution multiples as template, as shown in Figure 2, The DNA concentration of swimming lane 2 was 50ng / ul, and the ...

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Abstract

The invention belongs to the field of biotechnology and particularly provides a Periplaneta Americana L. specific COI primer, a kit containing the same and application of the Periplaneta Americana L. specific COI primer. According to a conserved sequence of Periplaneta Americana L. mtDNA, the invention designs a pair of the Periplaneta Americana L. specific COI primers JmF1 and JmR1 (such as SEQ ID No. 1 and SEQ ID No. 2), and detected that, the primer has the specific amplification ability to Periplaneta Americana L. only, the size of an amplified product is 452bp, and a sensitivity DNA concentration detection limit is 0.390625ng / uL. According to the Periplaneta Americana L. specific COI primer, the kit and the application, by utilizing a PCR detection technology, a specific single band is obtained through specific primer amplification, operation is simple, and the Periplaneta Americana L. specific COI primer has the characteristics of high efficiency, low cost and strong specificity and improves the accuracy of detection.

Description

technical field [0001] The invention belongs to the field of molecular biology, and specifically relates to a Periplaneta americana-specific COI primer, a kit containing the same and an application thereof. Background technique [0002] The American cockroach (Periplaneta Americana L.) is an insect of the genus Periplaneta in the subclass of Insecta Pterosae, Blattatae, Blattedae, commonly known as "cockroach", and is an important traditional medicinal material in my country. Its medicinal use was first recorded in "Shen Nong's Materia Medica", and it was listed as a middle-grade product. "Compendium of Materia Medica" records that its indications are "blood stasis, hard symptoms, cold and heat, lowering qi, and sharpening blood vessels." "Chinese Pharmacy Dictionary", "Chinese Medicine Dictionary", "Chinese Medicinal Animals", "National Chinese Herbal Medicine Compilation" and other recent and modern pharmacy monographs all record that cockroaches have "promoting blood cir...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/686C12Q1/6888C12Q2531/113
Inventor 姜顺日耿福能刘彬吴桃清
Owner SICHUAN GOODDOCTOR PANXI PHARMA
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