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A method for constructing a microsatellite multiplex PCR system of soft-shelled turtle genome

A technology of Chinese soft-shelled turtle and genome sequence, which is applied in the construction field of microsatellite marker multiplex PCR system, can solve the problems of lagging development and research of molecular markers, hindering the research progress of Chinese soft-shelled turtle genetics and breeding, and achieve rapid results, saving investment and reducing costs Effect

Active Publication Date: 2022-03-01
PEARL RIVER FISHERY RES INST CHINESE ACAD OF FISHERY SCI
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Problems solved by technology

At present, the artificial breeding of Chinese soft-shelled turtle has been carried out for many years, but there is a certain degree of lag in the development and research of molecular markers, which undoubtedly hinders the research process of Chinese soft-shelled turtle genetics and breeding.

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  • A method for constructing a microsatellite multiplex PCR system of soft-shelled turtle genome
  • A method for constructing a microsatellite multiplex PCR system of soft-shelled turtle genome
  • A method for constructing a microsatellite multiplex PCR system of soft-shelled turtle genome

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Embodiment Construction

[0023] The present invention is described in further detail below through specific examples. The examples are only used to illustrate the present invention, but not to limit the scope of the present invention. Unless otherwise specified, the examples are all in accordance with conventional experimental conditions or conditions suggested in the manufacturer's instructions. The reagents and instruments used in the examples were purchased from conventional biological product agents unless otherwise specified.

[0024] 1. Acquisition of Genome Information of Chinese Soft-shelled Soft-shelled Turtle

[0025] Use the public database to obtain the genome sequence of the Chinese soft-shelled turtle. The download address of the genome sequence of the Chinese soft-shelled turtle is ftp: / / ftp.ncbi.nlm.nih.gov / genomes / Pelodiscus sinensis / CHR Un / .

[0026] 2. Acquisition of microsatellite locus sequence information

[0027] MISA software was used to analyze the above-mentioned soft-s...

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Abstract

The invention discloses a technical method for rapidly constructing a multi-PCR system of soft-shelled soft-shelled turtle with 10× or more by using public data. Sequence analysis, using Primer 3 for batch primer design, using AutoDimer software to calculate the compatibility of primer sequences at polymorphic sites; 3) retaining the 10-12 sites with the strongest primer sequence compatibility to construct a multiplex PCR system, According to the concentrations of upstream primers 0.4 μM and downstream primers 10 μM for each site, prepare primer premixes, and then mix equal volumes of primer premixes for all sites to prepare multiplex PCR primer working solutions; 4) use conventional PCR methods for multiplex PCR systems Amplify and genotype the amplicons using capillary electrophoresis. The multiplex PCR system constructed by the invention has high labeling throughput, accurate and rapid genotyping results, and can provide molecular tools for research work such as the genetic breeding of soft-shelled turtles.

Description

technical field [0001] The invention relates to a method for constructing a microsatellite marker multiplex PCR system, in particular to a method for constructing a microsatellite marker multiplex PCR system based on the genome information of Chinese soft-shelled turtle. Background technique [0002] Microsatellite markers are co-dominant molecular markers, which consist of repeated repeating units (Motif, usually 1-6 bases) in the genomes of animals and plants and flanking sequences on both sides. Microsatellite markers have the characteristics of high variability, wide distribution in the genome, conforming to Mendelian inheritance, and easy PCR amplification. They are the most commonly used molecular markers in population genetics, paternity identification, and genetic management. [0003] The traditional genotyping method of microsatellite markers is to use polyacrylamide gel electrophoresis to separate the alleles of different fragment sizes in the PCR product, and to d...

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/686
CPCC12Q1/686C12Q2537/143C12Q2563/107
Inventor 陈辰朱新平李伟
Owner PEARL RIVER FISHERY RES INST CHINESE ACAD OF FISHERY SCI