Method for rapidly identifying mirror carps

A mirror carp, rapid technology, applied in biochemical equipment and methods, microbial determination/inspection, DNA/RNA fragments, etc., can solve problems such as pollution, inability to identify mirror carp genome, etc., to increase feasibility and accuracy. and reliability, improve work efficiency

Active Publication Date: 2015-12-30
CHINESE ACAD OF FISHERY SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Previous identification of mirror carp was mainly based on morphological aspects, however, morphological identification cannot identify whether mirror carp genome is polluted by other species of carp

Method used

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  • Method for rapidly identifying mirror carps
  • Method for rapidly identifying mirror carps
  • Method for rapidly identifying mirror carps

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] The gene sequences of mirror carp and other carp species were measured by next-generation sequencing technology. By analyzing the above-mentioned genome sequencing data and comparing the sequence differences of various carp strains, the missing sequence was found, as shown in SEQNO: 1; the primer design was directly in the deletion The upstream and downstream 200bp of the sequence are designed, and the designed primer sequences are shown in SEQNO:2 and SEQNO:3.

[0031] The above-mentioned primers provided by the present invention have high efficiency in PCR reaction and strong specificity for the amplification of the missing sequence.

Embodiment 2

[0033] 1) Take the live fish fins of mirror carp, purse red carp, colorful carp, Yellow River carp and Xingguo red carp, set 4 replicates for each fish, use NEB DNA extraction kit to extract the DNA of the fins of the above live fish, the specific operation See the instructions of the kit for steps.

[0034] 2) using the DNA of the above-mentioned mirror carp, purse red carp, colorful carp, Yellow River carp and Xingguo red carp as templates, and carrying out PCR amplification with the following primers (shown in SEQNO: 2 and SEQNO: 3),

[0035] Upstream primer 5′-TGCACGTTACTTTTAAACACTCC-3′

[0036] Downstream primer 5′-AAAAAGCAGCATGCCTTACAA-3′

[0037] The primers are designed by the inventors themselves and synthesized by Invitrogen. The PCR reaction system is a 15 μl reaction system, including: 10Xbuffer 1.5 μl; DNTP mixture 1.5 μl (2.5 μm / μl); DNA 2 μl; upstream primer (concentration 10 μm) 0.5 μl; Primer (10pm concentration) 0.5μl; sterile water 8.9μL.

[0038] The PCR...

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Abstract

The invention provides a method for rapidly identifying mirror carps. The method includes the following steps of 1, extracting tissue DNA; 2, carrying out PCR amplification; 3, carrying out electrophoresis detection. By means of the method, a small quantity of reagents and an experiment process rapid, convenient and easy to operate are applied, and only by cutting a small amount of pterygiophore or muscle, the mirror carps can be rapidly and accurately identified in a short time on the basis that the mirror carps are not killed; a practical production department and a scientific research department can more easily and rapidly identify the gotten samples, the accuracy and the reliability of the identification result are improved, and working efficiency is greatly improved.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for quickly identifying mirror carps. Background technique [0002] Mirror carp belongs to Osteichthyes, Cypriniformes, Cyprinidae. Mirror carp has the advantages of fast growth, high meat content, and good meat quality. It has been approved by the National Aquatic Variety Approval Committee as an excellent aquatic breeding species suitable for promotion in my country. Under reasonable stocking density and better feeding conditions, the mirror carp grows very fast. In the Heilongjiang area (production period 120 days), it can breed fish species with a size of 150 grams, and the second-year-old commercial fish can reach a size of more than 1 kg. . The feeding survival rate is about 98%, and the overwintering maturity rate is 96%. Under the condition that the stocking density of mirror carp is the same as that of other carps, the growth rate of German mirror carp exceeds th...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11C12Q1/68
Inventor 许建崔军徐鹏
Owner CHINESE ACAD OF FISHERY SCI
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