High-throughput single-cell transcriptome sequencing method and kit

Abstract

The invention discloses a high-throughput single-cell transcriptome sequencing method and a kit. The high-throughput single-cell transcriptome sequencing method includes the steps that a droplet generation system is adopted to encapsulate a single cell and labeled microbeads in a droplet, and reverse transcription is performed in the droplet. According to the high-throughput single-cell transcriptome sequencing method, a throughput of 9,000 cells which is equivalent to 10 x genomics can be achieved, after the droplet is demulsified, the microbeads are less contaminated with each other, and theproportion of valid data is increased. According to the high-throughput single-cell transcriptome sequencing method, reverse transcription is performed in the droplet, fewer reagents are required, and the cost is low. In the preferred scheme of the high-throughput single-cell transcriptome sequencing method, a SMART template conversion technology is adopted in reverse transcription, and productscan be directly used for subsequent Tn5 library construction and BGISeq-500 platform sequencing without library conversion; and multiple amplification and introduction of deviations are avoided, a droplet-based microfluidics platform is used in conjunction with a BGISeq-500 sequencing platform, and large-scale single-cell sequencing is simplified and facilitated.

Description

technical field [0001] This application relates to the field of single-cell sequencing, in particular to a high-throughput single-cell transcriptome sequencing method and kit. Background technique [0002] The mechanism of disease occurrence and their detection and prevention are mainly analyzed through the detection of cell genome variation and abnormal expression of transcriptome. Single-cell sequencing technology can reveal the complex heterogeneity of cells in tissues and provide accurate information for the diagnosis and treatment of diseases. Constrained by the processing throughput and cost of single-cell sequencing technology, many large-scale research work cannot be carried out. The combination of microfluidic technology and single-cell sequencing technology can solve these problems well; at the same time, single-molecule sequencing technology can avoid Errors caused by nucleic acid amplification have great application prospects and demands in basic scientific rese...

AI Technical Summary

Problems solved by technology

[0007] In general, the existing single-cell separation or single-cell transcriptome sequencing technologies are limited by issues such as throughput and cost, and many large-scale research work cannot be carried out

On the one hand, low throughput makes it difficult to achieve detailed analysis of cell heterogeneity and different cell subpopulations

On the other hand, the increased double packet rate after increasing the throughput has become a bottleneck of the current high-throughput platform

Images