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LAST-seq:single-cell RNA sequencing by direct amplification of single-stranded RNA without prior reverse transcription and second-strand synthesis

ABSTRACT: LAST-seq:single-cell RNA sequencing by direct amplification of single-stranded RNA without prior reverse transcription and second-strand synthesis

PROVIDER: PRJNA872200 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR21226621.fastq.gz	Fastqsanger.gz
	SRR21226622.fastq.gz	Fastqsanger.gz
	SRR21226623_1.fastq.gz	Fastqsanger.gz
	SRR21226623_2.fastq.gz	Fastqsanger.gz
	SRR21226624_1.fastq.gz	Fastqsanger.gz

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LAST-seq:single-cell RNA sequencing by direct amplification of single-stranded RNA without prior reverse transcription and second-strand synthesis

Project description:Existing single-cell RNA sequencing (scRNA-seq) methods rely on reverse transcription (RT) and second-strand synthesis (SSS) to convert single-stranded RNA into double-stranded DNA prior to amplification, with the limited RT/SSS efficiency compromising RNA detectability. Here, we develop a new scRNA-seq method, Linearly Amplified Single-stranded-RNA-derived Transcriptome sequencing (LAST-seq), which directly amplifies the original single-stranded RNA molecules without prior RT/SSS. LAST-seq offers a high single-molecule capture efficiency and a low level of technical noise for single-cell transcriptome analyses. Using LAST-seq, we characterize transcriptional bursting kinetics in human cells, revealing a role of topologically associating domains in transcription regulation.

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