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Triple-PCR (Polymerase Chain Reaction) detection method for exopalaemon carinicauda by using microsatellite mark

A technology of microsatellite marker and white shrimp, which is applied in the field of PCR detection of microsatellite markers of white shrimp, can solve the problems of missing and difficult evaluation of genetic parameters, and achieve the effects of easy detection, rich polymorphism, and shortened experiment time

Active Publication Date: 2012-09-19
YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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Problems solved by technology

[0005] Aiming at the shortcomings of lack of basic research work on white prawns in the prior art, which brings difficulties to the evaluation of genetic parameters in the process of family establishment, the present invention provides a 3-fold PCR detection method for microsatellite markers in white prawns. Based on a large number of microsatellite molecular markers, a 3-fold PCR detection system was constructed, which greatly shortened the experimental time and improved the experimental efficiency. The obtained results can simultaneously reflect the genetic variation of the white shrimp at multiple microsatellite loci

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  • Triple-PCR (Polymerase Chain Reaction) detection method for exopalaemon carinicauda by using microsatellite mark
  • Triple-PCR (Polymerase Chain Reaction) detection method for exopalaemon carinicauda by using microsatellite mark
  • Triple-PCR (Polymerase Chain Reaction) detection method for exopalaemon carinicauda by using microsatellite mark

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Embodiment 1

[0024] The 3-fold PCR construction method of the white shrimp microsatellite marker provided by the invention is as follows:

[0025] 1. Based on the microsatellite core sequence in the white shrimp microsatellite enrichment library provided by the present invention, use Primer Premier 6.0 software to design corresponding PCR primers at the two ends of its microsatellite core sequence to obtain a series of PCR primers with PCR amplification. Amplified SSR markers of varying product sizes.

[0026] 2. Use Primer Premier 6.0 software to evaluate between primers, and select the primers that show most of the ΔG (Gibbs of energy change) value above -4 for use, and the ΔG (Gibbs of energy change) value above -4 is mainly explained In the multiplex PCR system, it is difficult for a single pair of primers to form primer dimers and have a low hairpin structure, so as not to affect the efficiency of the PCR reaction. Through the collocation and combination of multiple pairs of microsat...

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Abstract

The invention provides a triple-PCR (Polymerase Chain Reaction) detection method for exopalaemon carinicauda by using a microsatellite mark. The triple-PCR detection method comprises the following steps of: designing and synthesizing a triple-PCR primer; extracting genome DNA (Deoxyribose Nucleic Acid); carrying out triple-PCR; and detecting a PCR product, wherein the PCR comprises a triple-PCR system and a triple-PCR program. According to the triple-PCR detection method disclosed by the invention, the triple-PCR system for identifying the family and the paternity of the exopalaemon carinicauda is established and synchronous detection of three microsatellite sites in one PCR is realized, so that the efficiency of the triple-PCR detection method is increased by three times compared with that of an original PCR method. The triple-PCR detection method disclosed by the invention has the characteristics of high efficiency, economy, simplicity, convenience, feasibility and the like and can be popularized and applied in genetic diversity analysis, genetic relationship identification, family management and selective breeding.

Description

technical field [0001] The invention relates to a PCR detection method for microsatellite markers of white shrimp, in particular to a triple PCR detection method for microsatellite markers of white shrimp. Background technique [0002] Microsatellites (microsatellites) or simple sequence repeats (simple sequence repeats, SSR) refers to a simple tandem repeat DNA sequence composed of 1 to 6 nucleotides. It has been found in all biological species studied so far, and its distribution density is very large. Because microsatellites are widely distributed in the genome, they have the characteristics of high density, rich polymorphism, high heterozygosity and good stability, co-dominant inheritance following Mendelian segregation law, and easy PCR amplification. It is an eye-catching new type of DNA marker, and is widely used in many research fields such as family genealogy authentication of biological resources, gene linkage analysis, genetic map construction, germplasm identifi...

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 刘萍李健贾舒雯
Owner YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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