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A molecular genetic identification method for channel catfish and longfin catfish

A technology for longfin catfish and channel catfish, applied in the field of genetics, can solve problems such as unguaranteed quality, mixed channel catfish and longfin catfish, and no literature reports on microsatellite repetition regularity, etc., to achieve high and stable repetition rate good sex effect

Active Publication Date: 2016-07-06
FRESHWATER FISHERIES RES INSITUTE OF JIANGSUPROVINCE +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the germplasm of channel catfish and longpin catfish depends on imports, and the quality cannot be guaranteed. There may be a mixture of channel catfish and longpin catfish
[0003] At present, there is no literature reporting the microsatellite duplication regularity of the channel catfish and long-fin catfish MSTN gene, and this is used as the basis for the molecular identification of the two fishes

Method used

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  • A molecular genetic identification method for channel catfish and longfin catfish
  • A molecular genetic identification method for channel catfish and longfin catfish

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] 1. Primer design

[0023] Design a pair of specific PCR amplification primers, the primer sequence is: upstream primer SEQIDNO.1: 5'-ACGGTGTTCCTGTTACTGC-3', downstream primer SEQIDNO.2: 5'-CACCAGATGTTGCTATGC-3'.

[0024] 2. Sample collection

[0025] Thirty channel catfish and 30 longfin catfish were collected.

[0026] 3. Genomic DNA extraction

[0027] Take about 20 mg of caudal fin tissue from each fish, blot the surface moisture with filter paper, put it into a 1.5 ml centrifuge tube, and add 420 μL STE buffer (30 mmol / LTris-HCl, pH8.0, 200 mmol / LEDTA, 50 mmol / L NaCl). After mixing, add 80 μL of 10% SDS and 10 μL of 20 mg / mL proteinase K, digest at 56°C for 8-10 hours; add an equal volume of phenol: chloroform: isoamyl alcohol (25:24:1) to extract twice, and centrifuge at 12000r After 10 minutes, take the supernatant; add an equal volume of chloroform:isoamyl alcohol (24:1) to extract once, centrifuge at 12000r for 10min, take the supernatant; add an equal volume...

Embodiment 2

[0034] According to the method in Example 1, the sample was expanded to 100 individual channel catfish and longfin catfish, and the results still showed that the number of (AGC) microsatellite repeats of channel catfish from the 83rd site (MSTN-83) was six times, and the length The number of (AGC) microsatellite repeats from the 83rd locus (MSTN-83) in the fin catfish was seven.

[0035]

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Abstract

The invention belongs to the genetics field and relates to a molecular genetics identification method for channel catfishes and flathead catfishes. The molecular genetics identification method for channel catfishes and flathead catfishes comprises the following steps: respectively extracting genome DNA (deoxyribonucleic acid) of to-be-identified channel catfishes and flathead catfishes; performing PCR (polymerase chain reaction) amplification on amicine genes of the channel catfishes and flathead catfishes; sequencing and comparing; if AGC microsatellites, starting from the 83th locus, of the MSTN (myostatin) gene are repeated for six times, identifying the catfishes as channel catfishes, and if the microsatellites are repeated for seven times, judging the catfishes as flathead catfishes. The molecular genetics identification method disclosed by the invention can be used for quickly, simply and conveniently, accurately and effectively identifying the channel catfishes and the flathead catfishes, is good in result stability and high in repetitive rate, so that the blank of identifying channel catfishes and flathead catfishes according to the existing domestic molecular biologic criteria is filled up.

Description

technical field [0001] The invention belongs to the field of genetics and relates to a molecular genetic identification method for channel catfish and longfin catfish. Background technique [0002] Channel catfish and longpin catfish are native to the United States. They are currently the species with the most mature breeding technology and the highest breeding output in the United States, with a breeding output of about 300,000 tons. Channel catfish has developed rapidly in just over 20 years since it was introduced into my country in the 1980s. It has successively broken through the technical difficulties in breeding, breeding, feed, processing and exporting, and has formed an industry from seed breeding to breeding. It has a complete industrial chain of production, processing, and marketing, and the output has also increased sharply from the initial few thousand tons to 224,000 tons at the peak, becoming one of the main varieties exported to the United States in my country...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6888C12Q2600/156
Inventor 钟立强陈校辉边文冀王明华秦钦陈友明张明生史阳白孔杰栾生
Owner FRESHWATER FISHERIES RES INSITUTE OF JIANGSUPROVINCE
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