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Primer and method for identifying acipenser baerii, amur sturgeon and cross generation thereof

A technology of Sturgeon's sturgeon and primer pairs, which is applied in the field of molecular biology and can solve problems such as complex sample preparation

Active Publication Date: 2018-10-16
SICHUAN AGRI UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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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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  • Primer and method for identifying acipenser baerii, amur sturgeon and cross generation thereof
  • Primer and method for identifying acipenser baerii, amur sturgeon and cross generation thereof

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Embodiment

[0063] (1) Using the sturgeon DNA to be identified as a template (the extraction method used the extraction method of the total genomic DNA in the previous 2), PCR amplification was performed using the microsatellite primer pair As1 as primers, and the total PCR reaction system was 25 μL (template 1 μL, Mix12. 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, extension at 72°C for 50 s; and finally extension at 72°C for 10 min. PCR reaction products were detected by 10% non-denaturing polyacrylamide gel (Polyacrylamide Gel, referred to as PAAG) electrophoresis, and after the electrophoresis was completed, a modified silver staining method was used for staining-development-scanning and imaging, and the data results were saved. If there is a 200bp amplification product but not a 105bp...

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Abstract

The invention discloses a primer and a method for identifying acipenser baerii, amur sturgeon and a cross generation thereof. The primer is composed of a acipenser baerii primer pair and a microsatellite primer pair As1, wherein a forward primer sequence of the acipenser baerii primer pair is as shown in SEQ ID No.1, and a backward primer sequence of the acipenser baerii primer pair is as shown inSEQ ID No.2; and a forward primer sequence of the microsatellite primer pair is as shown in SEQ ID No.3, and a backward primer sequence of the microsatellite primer pair is as shown in SEQ ID No.4. The primer and the method disclosed by the invention can be adopted to simply and quickly distinguish acipenser baerii, amur sturgeon and the cross generation thereof.

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 Applications(China)
IPC IPC(8): C12Q1/6888C12N15/11
CPCC12Q1/6888C12Q2600/156
Inventor 赵仲孟杨世勇陈虎黄小丽刘钊苗懿陈德芳杜宗君罗伟冯杨熊关庆段靖张瀚艺张谨啸朱凌威王云维陈益辉任妍李涛陈雨薇谭淦杨磊卢永瑞刘洪李
Owner SICHUAN AGRI UNIV
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