A method for exome assembly and sequencing

Abstract

The invention discloses an exon assembling and sequencing method, which relates to the technical field of exomes. The method comprises the steps of acquiring exon sequences dispersed at all parts of a gene, designing multiple pairs of primers in areas, close to the exons, on the gene, and amplifying the exon sequences through a PCR method; meanwhile, through a multiple PCR technology, enabling Tm values to be similar when designing the primers, so that the primers can be put in the same reaction system for amplifying at the same time; then on the basis of multiple PCR, adding a section of specific sequence-overlap between each two primers, amplifying among the exons, meanwhile, assembling through overlap, and forming a large amplicon containing a plurality of exon segements; carrying out one-time sequencing on the large amplicon so as to obtain a plurality of exon sequence information. According to the exon assembling and sequencing method provided by the invention, the exon sequencing operation can be carried out in a more effective way with lower cost.

Description

technical field [0001] The invention relates to the technical field of exomes, in particular to an exome assembly and sequencing method. Background technique [0002] The exome (exome) is the sum of all the exon regions in the genome of a species, and it is the most direct reflection of the function of the gene. By performing exome sequencing, genetic mutations associated with protein functional variation can be directly discovered. Since exons only account for 1% of the entire human gene sequence, and are extremely critical 1%, exome sequencing is more economical and efficient than whole genome resequencing. At present, exome sequencing technology has been applied to the research of finding pathogenic genes and susceptibility genes related to various complex diseases. [0003] Although exome sequencing technology is currently one of the important means of gene research, whether it is the traditional exon single capture sequencing method or the high-throughput sequencing t...

AI Technical Summary

Problems solved by technology

The next-generation sequencing technology fragments the genomic DNA, then connects adapters on both sides, and uses different methods to generate millions of space-fixed PCR monoclonal arrays. All single clones perform primer hybridization and enzyme The extension reaction, and taking pictures of the fluorescent label signals absorbed by each extension to obtain sequencing data, but the amount of data obtained is very large, and it is also a big challenge to analyze meaningful content in such a large amount of data. challenge

[0006] Existing gene exon sequencing methods basically design multiple pairs of primers to amplify each exon and then sequence them. However, the number of exons on human genes is relatively large, generally as many as 20-30. In this way, the separate amplification and sequencing of each exon is not only cumbersome and labor-intensive, but also has a large number of sequences and is expensive

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