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Molecular specific marker primer for identifying amaranthus viridis and two common adulterants and method thereof

A labeled primer and specific technology, which is applied in biochemical equipment and methods, microbiological determination/inspection, DNA/RNA fragments, etc., can solve the problem of easy destruction of medicinal materials, inability to identify by molecular marker technology, and low copy number of nuclear genome DNA And other issues

Active Publication Date: 2020-03-17
FUJIAN UNIV OF TRADITIONAL CHINESE MEDICINE
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, these genomic DNA-based molecular markers are susceptible to genomic DNA degradation, because nuclear genomic DNA has fewer copies than chloroplast DNA and is more likely to be destroyed in processed medicinal materials, making identification by its molecular markers impossible

Method used

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  • Molecular specific marker primer for identifying amaranthus viridis and two common adulterants and method thereof
  • Molecular specific marker primer for identifying amaranthus viridis and two common adulterants and method thereof
  • Molecular specific marker primer for identifying amaranthus viridis and two common adulterants and method thereof

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Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0026] 1 instrument

[0027] PCR instrument (Eppendorf, model 5332), electrophoresis system (Beijing Liuyi Instrument Factory, model DYY-12), low-temperature refrigerated centrifuge (Eppendorf, model 5810R), gel imaging analyzer (BIO-RAD ChemiDoc XRS), trace Pipettes (Eppendorf).

[0028] 2 reagents

[0029] 2×CTAB extract, 1×TAE buffer, agarose (Promega Company), ethidium bromide (Fluka Company), TransStart®TopTaq DNA Polymerase (Beijing Quanshijin Company), chloroform, absolute ethanol, iso All propanol was of domestic analytical grade.

[0030] 3 materials

[0031] The experimental samples in this study were identified and collected by Wei Yicong from Fujian University of Traditional Chinese Medicine and collected from different origins: 10 samples of Amaranthus unruffles, Amaranthus amaranthus and Amaranthus retroflexi were collected from different origins in China.

[0032] method

[0033] 4.1 Genomic DNA extraction:

[0034] ⑴ Take 5 g of fresh leaves of Amaranthus a...

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Abstract

The invention discloses a molecular specific marker primer for identifying amaranthus viridis and two common adulterants and a method thereof, which belong to the technical field of biology. Accordingto matK gene sequences of amaranthus viridis, amaranthus albus and amaranthus retroflexus, an upstream specific forward primer is designed: AvF:5'-CTTTTTGTTCCTATATAATCTAG-3', AaF: 5'-CTAGTAAAAGTCAAAGTTAAGCG-3', ArF: 5'-GACTCAATTCTTCAGTAATACCA-3', and a downstream universal reverse primer matk-R: 5'-TATAATAATGAGAAAGATTTCG-3'. A multiplex PCR system and a fluorescent quantitative PCR reaction system are respectively constructed by utilizing specific primers, so that rapid and accurate qualitative and quantitative identification among amaranthus viridis, amaranthus albus and amaranthus retroflexus is realized.

Description

technical field [0001] The invention belongs to the field of biological technology, and in particular relates to a molecular specific marker primer and a method for distinguishing amaranth and two common confounding products. Background technique [0002] The medicinal plant Amaranthus amaranthus ( Amaranthus viridis L.) The whole herb can be used as medicine, which has the effects of clearing away heat and detoxifying, diuretic and relieving pain. Amaranth is often used as a wild vegetable in folk because it is rich in protein, riboflavin, magnesium, fat, vitamin C, dietary fiber and other nutrients. Its wild plant germplasm resources are widely distributed, and cultivated varieties have not yet been formed. Due to the amaranth ( Amaranthus albus L. ), Amaranthus retroflexus ( Amaranthus retroflexus L.) is similar in shape to Amaranthus amaranthus, and has a similar distribution, often associated, and is easy to be confused and misused. Amaranth and amaranthus are ...

Claims

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Application Information

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IPC IPC(8): C12Q1/6895C12Q1/6858C12Q1/6851C12N15/11
CPCC12Q1/6895C12Q1/6858C12Q1/6851C12Q2600/156C12Q2600/16C12Q2531/113C12Q2537/143C12Q2563/107C12Q2537/165
Inventor 魏艺聪车苏容卢伟
Owner FUJIAN UNIV OF TRADITIONAL CHINESE MEDICINE
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