Rapid extraction method of shellfish deoxyribonucleic acid (DNA) for large-scale and high-throughput genotyping

Abstract

The invention provides a rapid extraction method of shellfish deoxyribonucleic acid (DNA) for large-scale and high-throughput genotyping. The method comprises the following steps of: firstly, selecting a little of gill tissue as a sampling tissue, then carrying out protein digestion and cell lysis to release DNA, and just centrifuging and absorbing an upper clear liquid and removing sediments. The upper clear liquid can be used for subsequent typing work of molecular markers after being diluted; the processes of protein extraction, DNA sedimentation and the like of a common DNA extraction method are removed; the entire process can be finished within an hour; the rapid extraction method has the advantages of rapid and simple operation, high quality of extracted DNA, long quality guarantee period and the like, and can be used for accurate typing of common molecular markers such as single nucleotide polymorphism (SNP) and the like. Just one or two gill filaments are selected, so that the effects on survival, growth, development and the like of shellfish are small; and the rapid extraction method is suitable for research work of a plurality of samples or precious samples, and is an excellent extraction method of rapid and effective non-fatal DNA for shellfish.

Description

Technical field [0001] The invention belongs to the technical field of molecular biology nucleic acid extraction, and specifically relates to a shellfish DNA rapid extraction method for large-scale, high-throughput genotyping. Background technique [0002] In the research of population genetics analysis and molecular breeding, high-throughput and large-scale genotyping of a large number of individuals is often required, and it is required not to kill precious organisms, and even sampling does not affect their normal growth. development. DNA is generally combined with proteins in cells. The extraction process of DNA is the process of breaking the cells, removing proteins, polysaccharides and lipids bound to the DNA, and finally separating and purifying the required DNA. For molecular biology research, the primary task of most of the work is to study the extraction of material DNA, and the acquisition of high-quality DNA has an important impact on the reliability of later data col...

AI Technical Summary

Problems solved by technology

However, these DNA extraction methods are generally cumbersome and time-consuming, require a large amount of starting tissue, cause great damage to the research individual, affect the subsequent growth and development of the organism, and even need to be killed. Therefore, these methods are not applicable For rapid sampling of living organisms or research where sampling cannot kill the research individual, such as molecular marker-assisted breeding

In mussels, non-lethal sampling methods have only been studied in mussels and oysters, mainly by clipping the mantle for tissue collection for subsequent genetic analysis, however recently it was found in mussels that the mantle Pruning causes higher mortality and shell deformities

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