Construction method of high-throughput single cell full-length transcriptome sequencing library, kit and sequencing method

Abstract

The invention discloses a construction method of a high-throughput single cell full-length transcriptome sequencing library, a kit and a sequencing method, and relates to the technical field of gene sequencing. According to the invention, high-throughput sequencing analysis of 10-107 single-cell full-length transcriptomes is realized simultaneously by creatively adopting a specific tag method on the basis of combining a droplet microfluidic technology, a splitting and combining technology and a high-throughput sequencing technology, and the technical problem of large-scale single-cell full-length transcriptome sequencing is solved; the existing single cell transcriptome sequencing technology is modified and perfected, the sequencing cost is reduced, and the detection flux is improved.

Description

technical field [0001] The invention relates to the technical field of gene sequencing, in particular, to a construction method and kit for a high-throughput single-cell full-length transcriptome sequencing library, and a sequencing method. Background technique [0002] Single-cell transcriptome sequencing (scRNA-seq) technology is a new focus in the global life sciences and a new tool for biomedical research. Single-cell transcriptome sequencing can sequence the entire RNA molecule at the single-cell level, which can reveal the heterogeneity between cells (Heterogeneity), not only can identify cell subtypes, but also reveal their functional status, developmental process, lineage Sources and interactions, etc., have extremely important application prospects in the fields of oncology, developmental biology, immunology, microbiology, and neuroscience. Therefore, various countries have launched a series of major single-cell funding programs, such as the Single Cell Analysis Pr...

AI Technical Summary

Problems solved by technology

[0006] At present, part of the manual operation of single-cell transcriptome sequencing has been replaced by automated equipment, but the overall cost is still high. Taking the Chromium system of 10X Genomics as an example, the preparation cost of a single cell is about 0.15-1 US dollars, which is not yet suitable for large-scale clinical application

[0007] Therefore, large-scale single-cell full-length transcriptome sequencing is still a major challenge for single-cell sequencing.

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