Method and kit for identifying six types of perciform fishes and raw meat products thereof based on DNA Barcoding

A technology for perciformes and identification methods, which is applied in biochemical equipment and methods, determination/inspection of microorganisms, etc., can solve the problems of long time, high cost, and cannot be used for screening and identification of fish, and achieves a simple analysis method. , the effect of saving sequencing costs

Inactive Publication Date: 2015-07-01
GANSU AGRI UNIV
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Problems solved by technology

However, because the traditional morphological taxonomy is not paid enough attention at present, the professionals of morphological taxonomy are gradually decreasing, which affects the work of fish resource protection and habitat research.
[0010] CN200510022323.1, a rapid identification method of fish molecular biology developed by Sichuan University in recent years, combines the conservation of nuclear DNA sequences encoding fish RNA and the interspecies differences of transcriptional spacers with the traditional Electrophoresis technology to identify fish, this method is time-consuming and costly, more importantly, this method cannot be used to screen and identify specific fish
There are no patent applications and reports on the identification of the above-mentioned perciformes based on DNA barcoding technology at home and abroad

Method used

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  • Method and kit for identifying six types of perciform fishes and raw meat products thereof based on DNA Barcoding
  • Method and kit for identifying six types of perciform fishes and raw meat products thereof based on DNA Barcoding
  • Method and kit for identifying six types of perciform fishes and raw meat products thereof based on DNA Barcoding

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Embodiment Construction

[0028] Method of the present invention, carry out according to the following steps:

[0029] 1. First log in GenBank (http: / / www.ncbi.nlm.nih.gov / ) in NCBI to browse the fish DNA barcode database, download Complete genome of fish, and use Primer Premier 5.0 software to design and synthesize primers.

[0030] 2. Then carry out the operation of fish DNA extraction, the operation steps are as follows:

[0031] (1) Take 10 mg of muscle tissue from each sample, and use ophthalmic anatomical scissors to cut it into pieces as much as possible.

[0032] (2) Carefully pack the extracted sample into a 1.5ml autoclaved centrifuge tube, and then add 600μl of SET (lysate).

[0033] (3) Add 10 μl of PROK (20) to each EP tube, then add 15 μl of SDS, and mix thoroughly.

[0034] (4) Digest overnight in a water bath at 55°C to fully digest the muscle tissue.

[0035] (5) Add an equal volume of about 600 μl of Tris-saturated phenol to each EP tube, shake and mix for 15 minutes, and then cent...

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Abstract

The invention discloses a method for identifying six perciform fishes and raw meat products thereof based on DNA Barcoding. The method comprises the following steps: (1) designing a synthetic primer according to the known fish DNA Barcoding database; (2) extracting various fish DNAs containing the six fishes, performing PCR amplification by the primer, and selecting the best primer and the reaction condition according to the amplified result; (3) performing amplification on the DNA of each to-be-tested sample by using the primer screened by the step (2), sequencing, correcting and comparing the amplified fragment to obtain the DNA Barcoding databases of various fishes; (4) detecting a new to-be-tested sample by using the primer screened by the step (2), comparing with the sequence length in the DNA Barcoding database of the step (3), and determining whether the to-be-tested sample is one of the six types of perciform fishes. The method provided by the invention is low in cost, simple and convenient in operation and high in accuracy; the identification for the six types of perciform fishes and the raw meat products thereof can be realized and a technical support for carrying out the works such as the food safety supervision and management and the species and environment protection can be provided.

Description

Background technique [0001] DNA barcoding is a technique for species identification by analyzing the DNA sequence of a standard target gene. The principle of this concept is the same as the product bar code used to identify products in the retail industry. Simply put, the key to DNA barcoding technology is to scan one or some related genes on a large scale to identify an unknown species or discover a new species (Hebert et al., 2003a; Hebert et al., 2003b; Hebert et al., 2003b; et al., 2004). [0002] Since the concept of DNA barcoding was proposed, this emerging taxonomic technique has attracted more and more biologists' attention. DNA barcoding technology is a new technology to assist species identification in taxonomy, and it represents a new direction of biological taxonomy research (Xiao Jinhua et al., 2004), so it will be widely used in many fields such as ecology, environment, food, etc. (Janzen, 2004). [0003] There are many main advantages of DNA barcoding techno...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6869C12Q2531/113C12Q2563/185
Inventor 蔡原赵生国权金强李永青汪文强郭永博郑王山边琳鹤
Owner GANSU AGRI UNIV
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