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Primer and method for molecular identification of channel catfish and ietalurus punetaus

A technology for channel catfish and cloud catfish, applied in the field of molecular biology, can solve problems such as escape, ecological damage in water areas, difficult identification, etc., and achieve the effects of high repetition rate, good stability, and wide application prospects

Active Publication Date: 2022-08-02
FRESHWATER FISHERIES RES INSITUTE OF JIANGSUPROVINCE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Channel catfish and cloud catfish are both scaleless Siluriformes fish, with similar shapes and the same habitat waters, especially in the juvenile stage, it is difficult to identify only by shape and color, and it is even more difficult to accurately make fish products identification
In addition, the extensive breeding of channel catfish and cloud catfish has caused certain escapes and caused certain damage to the ecology of our waters and native fish.

Method used

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  • Primer and method for molecular identification of channel catfish and ietalurus punetaus
  • Primer and method for molecular identification of channel catfish and ietalurus punetaus

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Example 1: Molecular identification of channel catfish, channel catfish and yellow catfish

[0023] 1. Primer design:

[0024] Design specific PCR amplification primer pairs, primer pair 1 sequence is: upstream primer SEQ ID NO.1: 5'-AAGACATTGGYACCCTYTAC-3', downstream primer SEQ ID NO.2: 5'-ATAGGAGGACRGCYGTRATA-3'.

[0025] 2. Sample collection

[0026] Channel catfish, channel catfish and yellow catfish to be identified, each with 30 individuals, were purchased from the market.

[0027] 3. Genomic DNA extraction

[0028] About 20 mg of caudal fin tissue was taken from each fish, and the genomic DNA of all individuals was extracted using a genome extraction kit. DNA integrity was identified by 1.5% agarose gel electrophoresis, and its OD value was measured by UV spectrophotometer, diluted to 50ng / μL, and stored at -20°C for later use.

[0029] 4. PCR amplification and detection

[0030] Using the extracted DNA as a template, PCR amplification was performed with th...

Embodiment 2

[0035] 1. Sample collection

[0036] According to the method of Example 1, the samples were expanded to 60 individuals of channel catfish, channel catfish and yellow catfish, all of which were purchased from the market.

[0037] 2. Total DNA extraction of samples

[0038] The total DNA of the fish samples to be tested was extracted using a genome extraction kit. The total DNA extraction method was the same as that in Example 1.

[0039] 3. PCR amplification and detection

[0040] The primers SEQ ID NO.1 and SEQ ID NO.2 were used for PCR amplification of the DNA to be detected and the total DNA, and the method was the same as that in Example 1. PCR products were detected and separated in 1% agarose gel electrophoresis. After electrophoresis at 120 V for 30 min, the size and purity of PCR amplification products were determined by gel imaging analysis system.

[0041] 4. Sequencing comparison

[0042] The detected PCR amplification product was directly sent to the biological...

Embodiment 3

[0044] Example 3 Molecular identification of channel catfish and channel catfish

[0045] 1. Primer design

[0046] A pair of specific PCR amplification primers were designed, the primer sequences are: upstream primer SEQ ID NO.3: 5'-AACCCGATTCTTCGCATTYC-3', downstream primer SEQ ID NO.4: 5'-CCGATGATGATRAATGGGTG-3'.

[0047] 2. Sample collection

[0048] Channel catfish, channel catfish and yellow catfish to be identified, 30 individuals each, were purchased from the market.

[0049] 3. Genomic DNA extraction

[0050] About 20 mg of caudal fin tissue was taken from each fish, and the genomic DNA of all individuals was extracted using a genome extraction kit. DNA integrity was identified by 1.5% agarose gel electrophoresis, and its OD value was measured by UV spectrophotometer, diluted to 50ng / μL, and stored at -20°C for later use.

[0051] 4. PCR amplification and detection

[0052] Using the extracted DNA as a template, PCR amplification was performed with the above prim...

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Abstract

The invention provides a primer and a method for molecular identification of channel catfish and leiocassis longirostris. The nucleotide sequence of the primer is shown as SEQ NO: 1 and SEQ NO: 2 or SEQ NO: 3 and SEQ NO: 4. Similar fishes such as channel catfish are identified from the cytochrome oxidase subunit I gene and the cytochrome b gene, and the two fishes can be rapidly, conveniently, accurately and reliably identified by utilizing the primers through a mitochondrial genome DNA bar code technology; the primer and the method can quickly, conveniently, accurately and reliably identify channel catfish, ietalurus punetaus and the like, are good in result stability and high in repetition rate, fill the blank of identifying channel catalurus punetaus, ietalurus punetaus and pelteobagrus fulvidraco according to domestic molecular biological standards at present, play an important role in ecological resource investigation and fish variety identification, and have good application prospects. Wide application prospects are realized.

Description

technical field [0001] The invention belongs to the technical field of molecular biology, and in particular relates to a primer and a method for molecular identification of channel catfish and channel catfish. Background technique [0002] channel catfish ( Ictalures punctatus ) and channel catfish ( Ameiurus nebulosus ) are all produced in North America and are large catfish. They were introduced into my country for trial breeding in the 1980s, and successively broke through the technical difficulties in breeding, breeding, feed, processing and exporting, and the output and breeding area continued to expand. It is one of the main cultured fishes of the order Catfish in my country. [0003] Channel catfish and channel catfish are both scaleless catfish with similar shapes and inhabit the same waters. Especially in the juvenile stage, it is difficult to identify only by shape and color, and it is even more difficult to accurately make fish products. identification. In add...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11C12Q1/6888C12Q1/6869
CPCC12Q1/6888C12Q1/6869C12Q2531/113C12Q2525/151Y02A40/81
Inventor 钟立强王明华陈校辉张世勇姜虎成
Owner FRESHWATER FISHERIES RES INSITUTE OF JIANGSUPROVINCE
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