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Smart-RRBS for single cell methylome and transcriptome analysis

ABSTRACT: Smart-RRBS for single cell methylome and transcriptome analysis

PROVIDER: PRJNA660164 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR12545800_1.fastq.gz	Fastqsanger.gz
	SRR12545800_2.fastq.gz	Fastqsanger.gz
	SRR12545900_1.fastq.gz	Fastqsanger.gz
	SRR12545900_2.fastq.gz	Fastqsanger.gz
	SRR12545801_1.fastq.gz	Fastqsanger.gz

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Project description:The integration of DNA methylation and transcriptional state within single cells is of broad interest. Several single-cell dual- and multi-omics approaches have been reported that enable further investigation into cellular heterogeneity, including the discovery and in-depth study of rare cell populations. Such analyses will continue to provide important mechanistic insights into the regulatory consequences of epigenetic modifications. We recently reported a new method for profiling the DNA methylome and transcriptome from the same single cells in a cancer research study. Here, we present details of the protocol and provide guidance on its utility. Our Smart-RRBS (reduced representation bisulfite sequencing) protocol combines Smart-seq2 and RRBS and entails physically separating mRNA from the genomic DNA. It generates paired epigenetic promoter and RNA-expression measurements for ~24% of protein-coding genes in a typical single cell. It also works for micro-dissected tissue samples comprising hundreds of cells. The protocol, excluding flow sorting of cells and sequencing, takes ~3 d to process up to 192 samples manually. It requires basic molecular biology expertise and laboratory equipment, including a PCR workstation with UV sterilization, a DNA fluorometer and a microfluidic electrophoresis system.

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