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Molecular specificity labeled primer for mutual discrimination of Amaranthus albus L., Amaranthus blitum L. and Amaranthus retroflexus L., and method

A technology of labeling primers and specificity, which is applied in biochemical equipment and methods, microbial determination/inspection, DNA/RNA fragments, etc. And other issues

Active Publication Date: 2020-04-03
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 specificity labeled primer for mutual discrimination of Amaranthus albus L., Amaranthus blitum L. and Amaranthus retroflexus L., and method
  • Molecular specificity labeled primer for mutual discrimination of Amaranthus albus L., Amaranthus blitum L. and Amaranthus retroflexus L., and method
  • Molecular specificity labeled primer for mutual discrimination of Amaranthus albus L., Amaranthus blitum L. and Amaranthus retroflexus L., and method

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Experimental program
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Effect test

Embodiment Construction

[0025] 1 instrument

[0026] PCR instrument (Eppendorf, model 5332), real-time quantitative PCR instrument (ABI7900), electrophoresis system (Beijing Liuyi Instrument Factory, model DYY-12), low-temperature refrigerated centrifuge (Eppendorf, model 5810R), gel imaging analyzer ( BIO-RAD ChemiDoc XRS), micropipette (Eppendorf).

[0027] 2 reagents

[0028] 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 is of domestic analytical grade.

[0029] 3 materials

[0030] The experimental samples in this study were identified and collected by Wei Yicong of Fujian University of Traditional Chinese Medicine and collected from different origins: 10 samples of green amaranth, amaranth and retroflexus were collected from different origins in China.

[0031] method

[0032] 4.1 Genomic DNA extraction:

[0033]⑴ Take 5 g of fresh green...

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Abstract

The invention discloses a molecular specificity labeled primer for mutual discrimination of Amaranthus albus L., Amaranthus blitum L. and Amaranthus retroflexus L., and a method, and belongs to the technical field of biology. According to matK gene sequences of the Amaranthus albus L., the Amaranthus blitum L. and the Amaranthus retroflexus L., upstream specific forward primers AalF: 5'-TTTCAAAAAGGAATCGAAGATAA-3', AblF: 5'-CGAATCCATTTTTACGGAAACA-3' and AreF: 5'-GGGACTCAATTCTTCAGTAATACCA-3', and a downstream universal reverse primer matKR: 5'-AATAATGAGAAAGATTTCGGCAT-3' are designed. The specific primers are used for respectively building a multiplex PCR system and a fluorescent quantitative PCR system, and the fast and accurate mutual qualitative and quantitative discrimination among the Amaranthus albus L., the Amaranthus blitum L. and the Amaranthus retroflexus L. is realized.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a molecular specific marker primer and a method for mutual identification of green amaranth, amaranthus amaranthus and reflexa reflexa. Background technique [0002] The medicinal plant Amaranth ( Amaranthus albus L. ) The whole plant can be used as medicine, also often called "white amaranth". It has the effect of clearing away heat and detoxification. External application can cure sores, gnats, and insect bites. Amaranth ( Amaranthus blitum L.) The whole herb is used as a medicine for relieving pain, astringent, diuretic and antipyretic. Amaranthus retroflexus ( Amaranthus retroflexus L.) is mainly used to treat diarrhea, dysentery, hemorrhoid swelling, pain and bleeding embolism. The three are similar in shape and have similar distributions, often associated with each other, and are easy to be confused and misused. But they have different properties and medi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895C12Q2600/156
Inventor 魏艺聪车苏容卢伟
Owner FUJIAN UNIV OF TRADITIONAL CHINESE MEDICINE
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