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Primer pair, kit and method for detecting the growth rate of puffer chrysanthemum

A growth rate, primer pair technology, applied in biochemical equipment and methods, microbial determination/inspection, DNA/RNA fragments, etc., can solve problems such as result interference, and achieve high accuracy and repeatability.

Active Publication Date: 2016-08-31
上海市水产研究所(上海市水产技术推广站)
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Problems solved by technology

[0006] On the basis of the above findings, in order to solve the problem of the interference of the PC amplification copy on the results in the prior art, and to make up for the gap in the research on the growth rate of the puffer chrysanthemum in the prior art

Method used

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  • Primer pair, kit and method for detecting the growth rate of puffer chrysanthemum
  • Primer pair, kit and method for detecting the growth rate of puffer chrysanthemum
  • Primer pair, kit and method for detecting the growth rate of puffer chrysanthemum

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Embodiment 1

[0023] Step 1. Selection of Fugu chrysanthemum with fast growth rate

[0024] Choose to get from half-year-old chrysanthemum yellow puffer, divide into two groups according to body weight and body length, adopt phenol chloroform extraction method (referring to Huang Peitang et al. September 2010) extracted the DNA from Fugu chrysanthemum, and designed and synthesized a pair of selective amplification primers.

[0025] Upstream primer 5'-CAAGCTTGTCTGTTGAGGCCTGTT-3',

[0026] Downstream primer 5'-CAGCACCTTCGACCTCATTTGATG-3';

[0027] PCR amplification of its DNA, reaction system 20μL, including 2ul10×PCR buffer, 0.8ul10mmol / L dNTP mix, 0.4μmol / L upstream primer, 0.4μmol / L downstream primer, 30ng sample Chrysanthemum yellow puffer For DNA, the reaction conditions are as follows: denature at 94°C for 3 minutes, then denature at 94°C for 30 seconds, anneal at 55°C for 30 seconds, and extend at 72°C for 30 seconds, 42 cycles, and finally extend at 72°C for 10 minutes.

[0028] Th...

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Abstract

The invention relates to a primer pair for detecting growth rate of takifugu flavidus, a kit and a method. The primer pair consists of an upstream primer with the base sequence of SEQ ID NO:3 and a downstream primer with the base sequence of SEQ ID NO:4. The primer pair is further used for preparing the kit. Besides the primer pair, the kit further comprises a takifugu flavidus MARKER in fast growth rate. The detection method comprises the following steps: extracting the DNA (Deoxyribonucleic Acid) of a sample takifugu flavidus, and carrying out PCR (Polymerase Chain Reaction) amplification on the DNA of the sample takifugu flavidus by the primers in the kit; and carrying out a PAGE treatment on an amplification product, observing the size of a stripe, wherein the individual which is consistent to the MARKER stripe is the one fast in growth rate while the individual which is inconsistent to the MARKER stripe is the one slow in micro growth rate. The growth rate of the takifugu flavidus is identified by means of the primer pair, the kit and the method provided by the invention, so that the invention has the advantages of high repeatability, strong accuracy, simplicity, convenience and fast speed.

Description

[technical field] [0001] The invention relates to detection primers and a method for puffer chrysanthemum, in particular to a pair of primers, a kit and a method for detecting the growth rate of puffer chrysanthemum. [Background technique] [0002] Fugu flavidus (Fugu flavidus), commonly known as Chuanba, Good Fish, Juhuang, Gypsophila, etc., belongs to Tetraodontif-ormes (Tetraodontif-ormes) Tetraodontidae (Tetraodontidae) and is mainly distributed in my country. Yellow Sea, East China Sea and Bohai Sea. Because of its delicious taste and rich nutrition, it is quite popular and is a famous and precious cultured species. In the breeding process, the standardization of the breeding environment is getting higher and higher. Individuals with fast growth can shorten the breeding time, improve the utilization rate of feed, and save the cost of breeding. Therefore, cultivating and screening fast-growing chrysanthemum yellow oriental puffer is the pursuit of aquaculture. Target. ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/686C12Q2531/113
Inventor 张之文张根玉施永海张海明张中华徐嘉波邓平平
Owner 上海市水产研究所(上海市水产技术推广站)
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