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Multi-PCR kit for rapidly identifying Dragonfish type and identifying method thereof

A kit and arowana technology, applied in the field of molecular biology identification, can solve the problems of disparity in form and ornamental value of adult fish, identification, difference in economic value, etc., and achieve the effects of saving experimental time, easy operation, and improved sensitivity

Inactive Publication Date: 2017-05-10
厦门海关技术中心
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the great differences in the adult fish form and ornamental value of these dragon fishes, their economic value also has huge differences, but the larval forms of these dragon fishes are very similar, and it is difficult to identify them by morphological methods

Method used

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  • Multi-PCR kit for rapidly identifying Dragonfish type and identifying method thereof
  • Multi-PCR kit for rapidly identifying Dragonfish type and identifying method thereof
  • Multi-PCR kit for rapidly identifying Dragonfish type and identifying method thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0046] Materials and Methods

[0047] 1. Materials

[0048] 1.1 Asian arowana, silver arowana and pearl arowana samples provided by farmers;

[0049] 1.2 Main instrument reagents

[0050]

[0051] 2. Method

[0052] 2.1 Mitochondrial DNA extraction

[0053] 1. Extract mitochondrial DNA from tissue, weigh 100mg sample, add 400μl lysate for homogenization until no obvious tissue pieces can be seen;

[0054] 2. Place in a 37°C water bath for 1 hour;

[0055] 3. Add 200μl 5mol / L sodium chloride solution, mix well and centrifuge at 13000rpm for 15min;

[0056] 4. Take the supernatant, extract twice with phenol and once with chloroform;

[0057] 5. Add twice the volume of absolute ethanol, 1 / 10 volume, 3mol / L, pH8.0 potassium acetate, store at -20°C for 1 hour,

[0058] 6. Centrifuge at 13000rpm for 15min, discard the supernatant, and wash the precipitate twice with 70% ethanol; 7. After drying at room temperature, dissolve it in 50μl TE solution (Shanghai Solaibao Biotech...

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Abstract

The invention provides a multi-PCR kit for rapidly identifying Dragonfish type and an identifying method thereof; the kit includes 1, three upstream primers and three downstream primers; 2, positive comparison product; 3, PCR reaction fluid; meanwhile, the method also provides a method for rapidly identifying the Dragonfish type, wherein the method includes steps of 1), selecting three upstream primers and three downstream primers; 2), preparing a gene group DNA template; 3), respectively adding the upstream primers and the downstream primers in the gene group DNA template prepared by the positive controlled DNA template and the Dragonfish to be tested, so as to perform PCR amplified reaction;4). Taking 5 miu L products after amplification and applying samples to 2.5% of sepharose gel; taking 50 bp Marker (stripes 50, 100, 150, 200, 250, 300, 350, 400, 450, 500) as standard molecule reference and performing electrophoresis; applying the electrophoresis result to the analysis of an ultraviolet gel imaging system; 5), judging result. The kit and the method can improve the identifying efficiency, save time, and save the identifying cost.

Description

technical field [0001] The invention belongs to the technical field of molecular biology identification, and in particular relates to a PCR kit for rapidly identifying arowana species and an identification method thereof. Background technique [0002] The dragon fish is graceful and flat, with a wide mouth and long whiskers. The torso is covered with neatly arranged and shining huge scales. When swimming, it is calm and majestic. It is highly sought after by ornamental fish lovers. It is the most valuable ornamental fish in the ornamental fish market. , especially in Chinese regions, the price of some arowanas exceeds one million yuan. [0003] Asian arowana (Scleropages formosus), pearl arowana (Scleropagesjardinii) and silver arowana (Osteoglossum bicirrhosum) are currently the most popular arowanas in the world. Asian Arowana and Pearl Arowana belong to the genus Osteoglossus, while Silver Arowana belongs to the genus Osteoglossus. Due to the great differences in the ad...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/686C12Q1/6888C12Q2600/16C12Q2600/166C12Q2537/143
Inventor 郭书林陈信忠龚艳清杨俊萍孔繁德吴山楠
Owner 厦门海关技术中心
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