High-throughput sequencing-based methods and kits for labeling and capturing one or more specific genes of multiple samples

Abstract

The invention relates to a marker based on high-throughput sequencing and a method and a kit for capturing one or multiple specific genes of multiple samples. The method includes steps of primary amplification, primer digestion, secondary amplification, mixing, recycling, sequencing and analyzing. In the method, a mixture of a universal sequence and specific primers of all genes is adopted for primary amplification, specific sequences of all genes act in multiple amplification at this moment, and the universal sequence is added in front of each gene; a tag sequence and the universal sequence are adopted for secondary amplification, the universal sequence acts in amplification at this moment, and the tag sequence which is recognizable is added in front of a final product. By introducing a unique primer tag sequence into each sample, when the samples are detected through second-generation high-throughput sequencing technology, a sequencing result of each sample can be retrieved through the corresponding unique primer tag sequence, and the method can be applied to detecting multiple different genetic loci of a lot of samples at the same time, so that sequencing cost is lowered greatly.

Description

technical field [0001] The present invention relates to the field of biotechnology, in particular to a method and a kit for labeling and capturing specific genes in multiple samples, in particular, to a method for labeling and capturing one or more specific genes in multiple samples based on high-throughput sequencing and kits. Background technique [0002] The emergence of a new generation of high-throughput sequencers has greatly reduced the cost of nucleic acid sequencing, which has the characteristics of high throughput, low cost, and low sequencing error rate. With the application of the second-generation high-throughput sequencing technology, the mixed nucleic acid molecules can be sequenced, and each independent sequence can be resolved and measured at the same time, making it possible for this sequencing technology to be developed in high-throughput markers. [0003] With the widespread development of high-throughput sequencing technology in human disease research a...

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to use high-throughput sequencing technology to simultaneously sequence and analyze specific gene-related sequences in a large number of samples, solve the problems of low throughput and high sequencing costs in traditional methods, and expand the application of second-generation sequencing technology in the field of PCR sequencing

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