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Primers and methods for identifying Acipenser schneideri, Acipenser siberia and their hybrid offspring

An identification method and technology of sturgeon, applied in the field of molecular biology, can solve problems such as complex sample preparation

Active Publication Date: 2021-10-29
SICHUAN AGRI UNIV
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  • Claims
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Problems solved by technology

Using this method for identification, although the operation is simple and easy to learn and master, but the sample preparation is complicated, the band difference between samples is small, and it is easily affected by the sample collection method and various conditions in the experimental process. Therefore, in recent years, There are few reports on the identification of sturgeons, and they are mostly used for the assessment and determination of the individual health of sturgeons under artificial feeding conditions.

Method used

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  • Primers and methods for identifying Acipenser schneideri, Acipenser siberia and their hybrid offspring
  • Primers and methods for identifying Acipenser schneideri, Acipenser siberia and their hybrid offspring

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Embodiment

[0060] Using the sturgeon DNA to be identified as a template (the extraction method used the extraction method of total genomic DNA in the previous 2), PCR amplification was performed with the microsatellite primer pair HLJS41 as primers, and the total PCR reaction system was 25 μL (template 1 μL, Mix 12.5 μL , ddHO 10.5 μL, upstream primer 0.5 μL, downstream primer 0.5 μL). The PCR reaction conditions were as follows: pre-denaturation at 95°C for 5 min; then 35 cycles, each cycle including denaturation at 94°C for 40 s, annealing at 55°C for 35 s, and extension at 72°C for 50 s; finally, extension at 72°C for 10 min. The PCR reaction products were detected by 10% non-denaturing polyacrylamide gel (Polyacrylamide Gel, referred to as PAAG) electrophoresis. After the electrophoresis, the improved silver staining method was used for staining-development-scanning imaging and the data results were saved. If the amplification product of 255 bp appears but not the amplification produ...

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Abstract

The invention discloses primers and a method for identifying Acipenser styloides, Siberian sturgeon and their hybrid offspring, which consist of a pair of primers for Acipenser schnecke and a pair of microsatellite primers HLJS41, wherein the sequence of the forward primer of the primers for Acipenser strenecki is shown as SEQ ID Shown in No.1, the backward primer sequence is shown in SEQ ID No.2; the forward primer sequence of the microsatellite primer pair HLJS41 is shown in SEQ ID No.3, and the backward primer sequence is shown in SEQ ID No.4 . By adopting the primers and the method of the present invention, Siberian sturgeon, Sturgeon's sturgeon and their hybrid progeny can be easily and quickly distinguished.

Description

technical field [0001] The invention belongs to the field of molecular biology, and in particular relates to primers and methods for identifying Siberian sturgeon, Sturgeon's sturgeon and their hybrid offspring. Background technique [0002] Sturgeon's sturgeon and Siberian sturgeon are two important cultured species in my country. Under natural conditions, Sturgeon's sturgeon is only distributed in the Heilongjiang (Amur River) water system. It is a unique species and an important economic fish in Heilongjiang. Its growth rate is fast, but its disease resistance is poor, it is not resistant to transportation, and it is too dependent on live bait It is difficult to domesticate and has a high mortality rate during the breeding process, which increases the risk of the sturgeon farming industry. The Siberian sturgeon is mainly distributed in Central Asia and Eastern Europe, and a small amount is distributed in the Irtysh River system in my country. Its growth rate is slow, but...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6888C12N15/11
CPCC12Q1/6888C12Q2600/156
Inventor 杨世勇赵仲孟黄小丽刘钊苗懿杜宗君罗伟陈德芳陈虎冯杨段靖熊关庆
Owner SICHUAN AGRI UNIV
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