General microsatellite multiplex fluorescent PCR method for hypophthalmichthys molitrix and hypophthalmichthys nobilis

Abstract

The invention relates to a general microsatellite multiplex fluorescent PCR method for hypophthalmichthys molitrix and hypophthalmichthys nobilis and belongs to the technical field of aquatic animal germplasm identification in the aquaculture field. The method comprises the steps: firstly, collecting hypophthalmichthys molitrix and hypophthalmichthys nobilis pterygiophore samples; utilizing a traditional phenol-chloroform method to extract genome DNA; according to primer pairs referred to by a target amplified fragment length difference requirement, choosing 3 groups of quadruple PCR systems with clear amplified bands; totally referring to 12 microsatellite markers; finally, performing fluorescence decoration and amplification verification on the primer. The multiplex PCR method disclosedby the invention has the characteristics of quickness in operation, convenience in implementation, low cost and the like and can provide a novel molecular marker assistive technology for researches ofhypophthalmichthys molitrix and hypophthalmichthys nobilis germplasm genetic evaluation, family breeding and the like.

Description

technical field [0001] The invention relates to a general-purpose microsatellite fluorescent multiplex PCR method for silver carp and bighead carp, which is suitable for research on genetic variation and parentage identification of silver carp and bighead carp populations, and belongs to the technical field of germplasm identification of aquatic animals in the field of aquaculture. Background technique [0002] Silver carp (Hypophthalmichthys molitrix) and bighead carp (Hypophthalmichthys nobilis) are closely related species in Cyprinidae. As important indigenous fishes in China, they are widely distributed in major rivers and lakes. Both are carnivorous fish, important biological resources of water purification fishery, and have important ecological value in current water environment protection and governance. At the same time, as the traditional bulk freshwater fish farming species in China, the two have a long history of farming and large breeding volume, occupying an ext...

AI Technical Summary

Problems solved by technology

In the past few decades, studies on the genetic diversity of wild resources of silver carp and bighead carp populations in China have also been carried out. Basic reference; there are also population genetic studies using microsatellite markers, but most of the detection techniques use polyacrylamide gel electrophoresis, and there is room for improvement and improvement in resolution and accuracy

However, microsatellite markers, which are commonly used molecular markers in animal pedigree identification, have not been reported in the paternity identification of silver carp and bighead carp; therefore, it is of great practical significance to screen suitable microsatellite markers and use them in the construction of paternity identification technology for silver carp and bighead carp

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