Construction method of single cell library

Abstract

The invention discloses a construction method of a single cell library. The method comprises the following steps: adopting a high-density solution as a suspension of single-cell labeled microbeads, and adopting a slender pipeline with a certain height as a liquid storage tank of the microbead suspension; the high-density solution greatly reduces the sedimentation rate of the large-specific-gravity labeled microbeads in a storage pool, and the purpose of maintaining uniform dispersion of the microbeads in the microfluidic process is achieved; and in the process of forming water-in-oil micro-droplets, the high-density solution is mixed with a cell / cell nucleus suspension in a certain ratio, and the solute of the high-density solution is diluted to a low concentration, so downstream library labeling and amplification reaction are carried out. According to the invention, the problem that the single-cell labeled microbeads are easy to settle in a microfluidic process to influence library preparation is solved; the scheme is suitable for marking microbeads with a series of densities and sizes, so a selection range is expanded. the method is simple in structure and convenient to operate, and a proper volume can be selected to avoid waste; and the characteristic of easy replacement is suitable for continuous preparation of a plurality of libraries.

Description

technical field [0001] The invention relates to the technical field of library construction, in particular to a method for constructing a single-cell library. Background technique [0002] Single-cell sequencing and omics analysis technologies play an important role in elucidating the heterogeneity of complex and diverse individual cells in tissues in terms of development and function. Among them, single-cell RNA sequencing can provide information on gene expression in individual cells; single-cell ATAC sequencing makes it possible to analyze epigenetic regulation at the single-cell level by capturing the chromatin opening state of individual cells. [0003] Water-in-oil micro-droplet technology based on microfluidic system is one of the important methods to realize single-cell sequencing. Micro-droplet encapsulation can realize single cell separation at the physical level; co-encapsulation of single cells and single labeled microbeads can realize the labeling of single-cel...

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Problems solved by technology

[0007] In view of the above deficiencies in the prior art, the purpose of the present invention is to provide a method for constructing a single-cell library, which aims to solve the existing problems of operational controllability and low degree of freedom, easily damaged microbeads, etc.

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