DNA barcode gene and identification method for high resolution fusion analysis and identification of true and false of walnut milk

A high-resolution melting and walnut milk technology, which is applied in the fields of high-resolution melting analysis to identify the authenticity of walnut milk, DNA barcode genes and identification, can solve the problems of walnut milk sequencing failure, sequencing failure, etc., and achieve good market application prospects, Wide range of applications, accurate and fast identification methods

Pending Publication Date: 2019-11-08
CHINA JILIANG UNIV
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Problems solved by technology

The application involves the use of ITS2DNA barcode genes for testing, but when determining the Jane's walnut kernel + peanut butter, the sequencing failed
The application is based on the amplified sequence of the ITS2 gene universal primer, which amplifies the entire ITS2 gene sequence of about 500 bp (sequence such as SEQ ID NO: 4), and found that there is an example of walnut milk sequencing failure

Method used

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  • DNA barcode gene and identification method for high resolution fusion analysis and identification of true and false of walnut milk
  • DNA barcode gene and identification method for high resolution fusion analysis and identification of true and false of walnut milk
  • DNA barcode gene and identification method for high resolution fusion analysis and identification of true and false of walnut milk

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Experimental program
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Embodiment Construction

[0028] 1. Screening of DNA barcodes of 10 kinds of nuts such as walnut and peanut

[0029] Due to the high fat and protein in nuts, it is difficult to extract DNA. DNA was extracted from walnuts, pecans, pecans, cashews, almonds, macadamia nuts, hazelnuts, peanuts, almonds, and pine nuts using the AxyGen kit method. The DNA concentration of the nuts mentioned is appropriate, OD260 / OD280 value standard, complete DNA bands can be seen in the DNA electrophoresis map, and the quality is good; CP-03 gene detection is used, and there are clear bands of appropriate size, which meet the conditions for amplification.

[0030] For 10 kinds of nuts including walnut and peanut, three kinds of DNA barcode genes, ITS2, psbA-trnH and RbcL, were used for PCR amplification. As shown in Table 1, obvious bands appeared in the electrophoresis results of PCR products, and the success rate of PCR amplification was 100%. The success rate of sequencing was higher than 90% for the other two genes exc...

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Abstract

The invention relates to the field of domain molecular identification, in particular to a DNA barcode gene and an identification method for high resolution fusion analysis and identification of true and false of walnut milk. According to the DNA bar code gene and the identification method, specific primer pairs ITS2-152 for walnuts are designed, and the ITS2 gene sequence of 152bp is successfullyamplified; and a miniature DNA bar code is used, an HRM method is adopted, and true and false identification of the walnut milk is realized.

Description

technical field [0001] The invention relates to the field of molecular identification, in particular to a DNA barcode gene and an identification method for identifying the authenticity of walnut milk by high-resolution melting analysis. Background technique [0002] Walnut milk not only has the characteristics of pure natural plant protein raw materials, without losing the original nutritional content of walnut kernels and enhancing brain memory function, but also a good source of protein, vitamin B, niacin and various trace elements. It has always been a best-selling product in my country [1-2] . However, adulteration of walnut milk has caused false phenomena to emerge in an endless stream, and the means and methods are constantly being refurbished. It is difficult to detect and identify. The unknown allergen ingredients pose a threat to the personal health of consumers and affect food safety and trade order. [0003] At present, the identification of food authenticity mai...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12Q1/6851C12N15/11
CPCC12Q1/6895C12Q1/6851C12Q2531/113C12Q2527/107C12Q2563/107
Inventor 丁艳菲黄丽红朱诚姜广泽孙骏威李喜凤江琼
Owner CHINA JILIANG UNIV
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