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Method, special primer set and reagent kit for multiplex PCR identification of counterfeit product mixing of pine pollen

A technology of pine pollen and primer set, which is applied in the direction of recombinant DNA technology, biochemical equipment and methods, and the measurement/testing of microorganisms, etc. It can solve the problems of inability to meet the rapid detection of a large number of samples, the high requirements of experimental instruments, and the cumbersome and complicated experimental operations, etc. problems, to achieve the effect of standardizing the market order of pine pollen and ensuring product quality

Active Publication Date: 2019-12-13
YANTAI NEW ERA HEALTH IND
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Problems solved by technology

However, chemical composition analysis and fingerprinting require a series of complex operations such as pretreatment of pine pollen and extraction of characteristic compounds, which also have high requirements for experimental instruments.
DNA barcoding technology can identify pine pollen and its counterfeit products, but this method needs to amplify and sequence the barcode sequence of the sample every time it is measured, and needs to build a neighbor-joining tree. The experimental operation is cumbersome and complicated, and it cannot satisfy a large number of samples. rapid detection of

Method used

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  • Method, special primer set and reagent kit for multiplex PCR identification of counterfeit product mixing of pine pollen
  • Method, special primer set and reagent kit for multiplex PCR identification of counterfeit product mixing of pine pollen
  • Method, special primer set and reagent kit for multiplex PCR identification of counterfeit product mixing of pine pollen

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Embodiment 1

[0028] Primer design was carried out, and the primer set of the present invention was used for multiplex PCR identification of pine pollen adulteration.

[0029] 1. Design of multiple PCR primers for adulteration detection of pine pollen

[0030] According to the specific sequence of chloroplast trnL-F gene of pine pollen, Puhuang and maize, the multiplex PCR special primers for adulteration detection of pine pollen were designed.

[0031] 1. Amplification and alignment of chloroplast trnL-F gene sequences of pine pollen, Puhuang and maize

[0032] 1.1 Genomic DNA extracted from pine pollen, catarrh, and corn

[0033] Take an appropriate amount of pine pollen (collected in Yunnan, Chongqing, Hunan, Hebei, Shandong), Puhuang (purchased from a family of traditional Chinese medicine), corn flour (purchased from the market), grind it into a fine powder with liquid nitrogen, and extract it with plant genomic DNA Kits (DNeasy Plant MiniKit, QIAGEN) were used to extract genomic DNA...

Embodiment 2

[0047] A kit for multiplex PCR identification of pine pollen adulteration, the contents of which are as follows:

[0048] The specific primer pyF of the 0.33 μ M pine pollen chloroplast trnL-F gene, the 0.33 μ M of the yellow chloroplast trnL-F gene primer typF, the maize primer zmF of 0.67 μ M, and the universal reverse primer utR of 0.67 μ M are packaged together to obtain A multiplex PCR identification kit for pine pollen adulteration in a 30 μL reaction system.

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Abstract

The invention provides a special primer set for multiplex PCR identification of counterfeit product mixing of pine pollen. The special primer set comprises 3 specific forward primers designed in accordance with chloroplast trnL-F genes of pine pollen, cattail pollen and corn flour, and a reverse primer universal for the 3 chloroplast trnL-F genes. Through the adoption of the special primer set disclosed by the invention, the problem that raw materials in pine pollen market are mixed with counterfeit products can be solved, an objective criterion is provided for identifying the raw material sources of the pine pollen, and the special primer set has important significance in guaranteeing the product quality of the pine pollen and standardizing the order of the pine pollen market.

Description

technical field [0001] The invention relates to a multiplex PCR identification method for pine pollen adulteration and its special primer set and kit. Background technique [0002] Pine pollen, also known as pine flower and pine yellow, refers to the dried pollen of Pinus massoniana, Yunnan pine, Chinese pine, black pine, red pine or several plants of the same genus. It is a pollen variety used in traditional food and medicine in my country. Pine pollen is sweet and flat in nature and taste, returns to the liver and spleen meridians, and has the effects of nourishing the heart, lungs and Qi, strengthening the face and benefiting the will. From ancient times to the present, more than 30 important traditional Chinese medicine works in China have detailed records. Pine pollen contains free amino acids, polysaccharides, vitamins, flavonoids, trace elements and other components. It has rich nutritional value and various biological activities. It has broad prospects in both clini...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12Q1/686C12N15/11
CPCC12Q1/6895C12Q1/686C12Q2600/16C12Q2537/143C12Q2565/125
Inventor 仲米存衣洁菡尹雷王桐王洪涛郝娜曲涛
Owner YANTAI NEW ERA HEALTH IND
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