Amplicon sequencing method based on DNBSEQ platform and application thereof

Abstract

The invention provides an amplicon sequencing method based on a DNBSEQ platform, an amplicon library construction primer, an amplicon library construction method and a kit thereof. Specifically, the method comprises the following steps: constructing an amplicon library by adopting a primer with a special structure; loading the DNB to a regular array chip (normally loaded) for the second time; during sequencing, only sequencing template base biochemical synthesis is carried out on amplicons corresponding to upstream and downstream amplification primer sequences of a target region in first chain sequencing and second chain sequencing, and optical information collection is not carried out. The library and the sequencing analysis scheme adaptive to the DNBSEQ sequencing platform are efficiently and rapidly constructed through a one-step method, the application of amplicon products in the DNBSEQ platform is achieved, the method can simplify the design of library building primers, reduce the library building cost and simplify the data analysis process, and the method is suitable for large-scale popularization and application. The problem of randomness of basic groups in a target region can be effectively improved, so that the sequencing quality of an amplicon region is improved.

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