Molecular specific primer and method for identifying catfish

A technology for labeling primers and specificity, which is used in biochemical equipment and methods, microbial determination/inspection, DNA/RNA fragments, etc. It can solve problems such as lack of data, and achieve the effect of simple method and high sensitivity

Inactive Publication Date: 2018-03-27
ZHENJIANG HUADA DETECTION CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, due to the endemism of many fishes in

Method used

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  • Molecular specific primer and method for identifying catfish
  • Molecular specific primer and method for identifying catfish

Examples

Experimental program
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Effect test

Embodiment 1

[0034] (1) Extraction of genomic DNA of catfish to be tested

[0035] A catfish tissue sample was taken, cut into pieces, and extracted with a genomic DNA extraction kit. The DNA was detected by electrophoresis using 1.5% agarose gel, and the DNA concentration was detected by an ultraviolet spectrophotometer, and stored in a refrigerator at -20°C for use.

[0036] (2) Design and synthesize specific PCR amplification primers

[0037] Design and synthesis of catfish-based specific primers, upstream primer NYF: 5'-TTGCCCACGCAGGAGCT-3' and downstream primer NYR: 5'-CTGTAATTAGTACGGCTCAT-3', designed by technicians and synthesized by primer design company.

[0038] (3) PCR amplification of molecular specific marker primers (NYF / NYR)

[0039] PCR reaction system 25μL: 10×PCR Buffer 2.5μL, dNTPS 0.5μL, Taq(5U / μl) 0.2μL, MgCl 2 1.5μL, NYF primer (10μM) 1μL, NYR primer (10μM) 1μL, DNA template (about 10ng / μl) 1μL, ddH 2 O17.3μL.

[0040] The PCR reaction program is: 94°C pre-denaturation for 5 min...

Embodiment 2

[0045] The method of Example 1 was used to detect three kinds of frozen fish pieces of catfish, yellow catfish, and silver carp. The sampler made a record and numbered the species according to the specific information. The experiment was carried out when the experimenter did not know the name of the species. The result showed that there was only one Seed fish pieces can amplify specific DNA bands with a molecular weight of about 170 bp. The other two fish pieces samples cannot be amplified. The detection results are completely consistent with the information recorded by the sampler. Therefore, it can be shown that the nucleic acid molecule primer of the present invention has extremely high specificity, and therefore can be used for rapid identification of catfish.

Embodiment 3

[0047] Two groups of muscle tissue samples were selected from fish tissue samples and stored in 95% alcohol. The selected muscle tissue belongs to two groups of fish of different species, one of which is catfish (5 samples), and the other is other species (5 samples). The sampler will record and number the species according to specific information. The operator conducts the experiment without knowing the species belonging to the two groups of samples. The experiment was carried out using the same experimental method as in Example 1. The PCR product sequencing results showed that the 5 samples in one group could amplify the specific DNA bands of about 170 bp, which were catfish tissue samples, and the 5 samples in the other group were For tissue samples of non-catfish, the experimental results are completely consistent with the information recorded by the sampler.

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Abstract

The invention discloses a molecular specific primer and a method for identifying catfish. The molecular specific primer for identifying catfish has a forward primer NYF shown as SEQ ID NO: 1, and a reverse primer NYR shown as SEQ ID NO: 2. By utilizing the molecular specific labeled primer NYF/NYR, catfish can be subjected to rapid molecular identification by a conventional PCR method. The methodis simple, is capable of finishing detection with PCR technology and is an effective assistance for identifying catfish through morphological characteristics.

Description

Technical field [0001] The invention relates to a molecular specific marker primer for identifying catfish, and a method for quickly identifying catfish specimens by using the specific primer. Background technique [0002] Catfish (Scientific name: Silurus asotus) catfish, animal kingdom-Chordate phylum-Chordate subphylum-Radical fishes-Siluriformes-Catfish family-Catfish genus-Catfish species. The same species of catfish are distributed almost all over the world. Most species live in freshwater such as ponds or rivers, but some species live in the ocean. The general body has no scales, with a flat head and a large mouth. There are several long whiskers around the mouth. The taste can be distinguished by this whisker. This is its characteristic. At present, the species identification of catfish is limited to morphological identification, and there are few professional identification personnel. The construction of dams in the upper reaches of the Yangtze River resulted in the de...

Claims

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Application Information

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IPC IPC(8): C12Q1/6888C12N15/11
CPCC12Q1/6888
Inventor 崔海月张继敏蒋寿佳游欣欣石琼马兴宇
Owner ZHENJIANG HUADA DETECTION CO LTD
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