A rapid identification method of mirror carp

A mirror carp, rapid technology, applied in the direction of DNA / RNA fragments, recombinant DNA technology, etc., can solve the problems of pollution and inability to identify the mirror carp genome, etc., to increase feasibility, increase accuracy and reliability, and improve work efficiency Effect

A mirror carp, rapid technology, applied in the direction of DNA / RNA fragments, recombinant DNA technology, etc., can solve the problems of pollution and inability to identify the mirror carp genome, etc., to increase feasibility, increase accuracy and reliability, and improve work efficiency Effect

CN105200042BActive Publication Date: 2017-11-28FISHERY ENG RES INST CHINESE ACAD OF FISHERY SCI
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  • A rapid identification method of mirror carp
  • A rapid identification method of mirror carp
  • A rapid identification method of mirror carp

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 genome sequencing data and comparing the sequence differences of carp strains, the missing sequence was found, as shown in SEQ NO: 1; the primer design was directly in The upstream and downstream 200bp of the deletion sequence are designed, and the designed primer sequences are shown in SEQ NO:2 and SEQ NO: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 SEQ NO: 2 and SEQ NO: 3),

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

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

[0037] The primers were designed by the inventors themselves and synthesized by Invitrogen. The PCR reaction system was a 15 μl reaction system, including: 1.5 μl of 10X buffer; 1.5 μl of DNTP mixture (2.5 μm / μl); 2 μl of DNA; 0.5 μl of upstream primer (10 μm concentration) ; downstream primer (concentration 10pm) 0.5μl; sterile w...

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Abstract

The invention provides a method for rapidly identifying mirror carp, which comprises the following steps: 1) extracting tissue DNA; 2) PCR amplification; 3) electrophoresis detection. This method uses less reagents and a convenient, quick and easy-to-operate experimental process. It can quickly and accurately identify mirror carp in a short period of time without killing the fish species and only needs to cut off a small amount of fin rays or muscles. . This method is more beneficial to the actual production department and the scientific research department to quickly identify the samples taken, which increases the accuracy and credibility of the identification results and greatly improves the work efficiency.

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

Patent Timeline
28 Nov 2017
Publication
CN105200042B
IPC
C12N15/11; C12Q1/68
Inventors
许建; 崔军