A Highly Sensitive, Accurate, and Automated Single-Cell RNA Sequencing Platform with Digital Microfluidics

Multicellular organisms have many cell types and are complex, and heterogeneity is common among cells. Single-Cell RNA Sequencing (scRNA-SEQ) is a new technique for studying the transcriptional activity of a single cell that is still in its early stages of development. It generates transcriptional profiles from thousands of parallel cells to reveal the differential expression of individual cell genomes. They reflect the heterogeneity between cells to identify different cell types and form cell maps of tissues or organs, which play an essential role in biology and clinical medicine. Based on the introduction and comparison of the scRNA-SEQ sequencing platform, this paper focuses on the application of scRNA-SEQ in the exploration of cell types in the nervous system and immune system and summarizes the research results of the combination of scRNA-SEQ and spatial transcriptome technology.

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Lightning Fast and Highly Sensitive Full-Length Single-cell sequencing using FLASH-Seq

10.21203/rs.3.rs-715504/v1 ◽

2021 ◽

Author(s):

Vincent Hahaut ◽

Dinko Pavlinic ◽

Cameron Cowan ◽

Simone Picelli

Keyword(s):

Single Cell ◽

Rna Sequencing ◽

Exponential Growth ◽

Full Length ◽

Cdna Libraries ◽

Single Cell Sequencing ◽

Emulsion Droplets ◽

Highly Sensitive ◽

Single Cell Rna Sequencing ◽

Processing Steps

Abstract In the last 10 years, single-cell RNA-sequencing (scRNA-seq) has undergone exponential growth. Emulsion droplets methods, such as those commercialized by 10x Genomics, have allowed researchers to analyze tens of thousands of cells in parallel in a robust and reproducible way. However, in contrast to SMART-based full-length sequencing protocols, these methods interrogate only the outer portion of the transcripts and still lack the required sensitivity for analyzing comprehensively the transcriptome of individual cells. Building upon the existing SMART-seq forerunners protocols, we developed FLASH-Seq (FS), a new scRNA-seq method which displays greater sensitivity while decreasing incubation times and reducing the number of processing steps compared to its predecessors. The entire FS protocol - from lysed cells to pooled cDNA libraries - can be performed in ~4.5 hours, is automation-friendly and can be easily miniaturized to decrease costs.

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Lightning Fast and Highly Sensitive Full-Length Single-cell sequencing using FLASH-Seq

10.1101/2021.07.14.452217 ◽

2021 ◽

Author(s):

Vincent Hahaut ◽

Dinko Pavlinic ◽

Cameron S Cowan ◽

Simone Picelli

Keyword(s):

Single Cell ◽

Rna Sequencing ◽

Exponential Growth ◽

Full Length ◽

Cdna Libraries ◽

Single Cell Sequencing ◽

Emulsion Droplets ◽

Highly Sensitive ◽

Single Cell Rna Sequencing ◽

Processing Steps

In the last 10 years, single-cell RNA-sequencing (scRNA-seq) has undergone exponential growth. Emulsion droplets methods, such as those commercialized by 10x Genomics, have allowed researchers to analyze tens of thousands of cells in parallel in a robust and reproducible way. However, in contrast to SMART-based full-length sequencing protocols, these methods interrogate only the outer portion of the transcripts and still lack the required sensitivity for analyzing comprehensively the transcriptome of individual cells. Building upon the existing SMART-seq forerunners protocols, we developed FLASH-Seq (FS), a new scRNA-seq method which displays greater sensitivity while decreasing incubation times and reducing the number of processing steps compared to its predecessors. The entire FS protocol - from lysed cells to pooled cDNA libraries - can be performed in ~4.5 hours, is automation-friendly and can be easily miniaturized to decrease costs.

Download Full-text