Method and kit for enriching 4000 human pathogenic target genes

Abstract

The invention relates to a method and a kit for enriching 4000 human pathogenic target genes. The method comprises the following steps: breaking DNA to obtain DNA fragments, carrying out terminal repairing, carrying out A addition, carrying out linker addition, and carrying out Pre-Capture LM-PCR; mixing Pre-Capture LM-PCR products of 4000 human pathogenic target genes to obtain a mixed library sample, hybridizing a specific probe library of the 4000 human pathogenic target genes with the mixed library sample, capturing DNA by adopting magnetic beads, and carrying out eluting; and carrying out Post-Capture LM-PCR. The kit contains a DNA terminal repairing reagent, an A addition reagent, a linker addition reagent, an LM-PCR reagent and a capturing reagent. According to the method and the kit, the 4000 human pathogenic genes are taken as target spots and are combined together, the gene sequences are enriched once through a biotin probe capturing technology and are subjected to high-throughput sequencing, therefore, multiple variation types of the genes can be simultaneously examined, and the detection sensitivity is high; the 4000 pathogenic genes can be taken as a principal diagnosis and detection means clinically, the cost is reduced, the missed diagnosis is reduced, the detection positive rate is improved, and therefore, the clinical popularization is easy.

Description

technical field [0001] The invention relates to the field of gene detection, in particular to a method and kit for enriching 4000 human disease-causing target genes. Background technique [0002] The human genome is composed of 23 pairs of chromosomes, containing about 3 billion DNA base pairs. The euchromatin gene sequence in the human genome surveyed in the Human Genome Project contains approximately 20,000 to 25,000 protein-coding genes. Protein-coding sequences (ie, exons) make up less than 1.5% of the human genome. Beyond the gene and regulatory sequences, there are still many vast regions of unknown function, including many repeat sequences, transposons or pseudogenes. When one or more genes are abnormally expressed, it may cause a corresponding phenotype to produce some clinical symptoms. Causes of genetic abnormalities include gene mutations and abnormalities in the number of chromosomes. If damaged genes are passed from parent to offspring, it becomes a genetic ...

AI Technical Summary

Problems solved by technology

However, although next-generation sequencing has greatly reduced the cost of DNA analysis, its sample preparation process is still cumbersome, which has become an obstacle to its widespread application in the future

Images