Mass acquisition method of different loci and flanking sequence thereof in genome DNAs (deoxyribonucleic acids)

Abstract

The invention discloses a mass acquisition method of different loci and flanking sequence thereof in genome DNAs (deoxyribonucleic acids). The method comprises the following steps: mixing a genome DNA A and a genome DNA B, hybridizing, digesting the single chain DNA area which has not form a double chain area by using CELI endonuclease, recovering the DNA segments within the target size range in the digestion production, and carrying out self-hybridization; and carrying out DNA terminal filling-in, linker DNA addition, PCR (polymerase chain reaction) amplification and sequencing to finally obtaining the different loci in the genome DNAs A and B. The method disclosed by the invention has the advantages of high efficiency, high flux and low cost, and one positive clone can provide information of 2 genome DNA different loci; and the method is simple and convenient to operate, does not need complex equipment or expensive reagents before sequencing, and can be widely used for detecting and analyzing genome DNA different loci among species and strains of animals, plants and microbes, and between cancer cells and normal cells.

Description

technical field [0001] The invention relates to a method for obtaining differential sites and flanking sequences in genome DNA in batches. Background technique [0002] In the process of species evolution, the genome differences between different species are mostly manifested in different genome sizes, chromosome rearrangements, gene structure changes, base insertions, base deletions, and a large number of single-base nucleotide mutations (SNP) ; Genome differences between different individuals of the same variety (line) are mainly manifested as insertions, deletions of several bases, and a large number of SNPs. These different forms of genome differences or mutations cause large phenotypic differences among different individuals, and mutations of some bases can even cause the occurrence of diseases and the termination of development of living organisms. The acquisition of genomic difference loci is of great significance for revealing the reasons for different phenotypes am...

AI Technical Summary

Problems solved by technology

Among them, although high-throughput sequencing technology and chip technology have a large analysis throughput, they are expensive and require special equipment and reagents, while the analysis throughput of other technologies is too small to be widely used in genomic differential analysis

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