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Comparative analysis of single-cell RNA sequencing methods with and without sample multiplexing

ABSTRACT: Comparative analysis of single-cell RNA sequencing methods with and without sample multiplexing

PROVIDER: PRJNA1033995 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR26594138_1.fastq.gz	Fastqsanger.gz
	SRR26594138_2.fastq.gz	Fastqsanger.gz
	SRR26594139_1.fastq.gz	Fastqsanger.gz
	SRR26594139_2.fastq.gz	Fastqsanger.gz
	SRR26594140_1.fastq.gz	Fastqsanger.gz

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Comparative analysis of single-cell RNA sequencing methods with and without sample multiplexing

Project description:Single-cell RNA sequencing (scRNA-seq) has emerged as a powerful technique for investigating biological heterogeneity at the single-cell level in human systems and model organisms. Recent advances in scRNA-seq have enabled the pooling of cells from multiple samples into single libraries, thereby increasing sample throughput while reducing technical batch effects, library preparation time, and the overall cost. However, a comparative analysis of scRNA-seq methods with and without sample multiplexing is lacking. In this study, we benchmarked methods from two representative platforms: Parse Biosciences (Parse; with sample multiplexing) and 10X Genomics (10x; without sample multiplexing). By using peripheral blood mononuclear cells (PBMCs) obtained from two healthy individuals, we demonstrate that demultiplexed scRNA-seq data obtained from Parse showed similar cell type frequencies compared to 10X data where samples are not multiplexed. Despite a relatively lower library and cell capture efficiencies, Parse can detect rare cell types (e.g., plasmablasts and dendritic cells) which is likely due to its relatively higher sensitivity in gene detection. Moreover, comparative analysis of transcript quantification between the two platforms revealed platform-specific distributions of gene length and GC content. These results offer guidance for researchers in designing high-throughput scRNA-seq studies.

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Project description:These datasets are test datasets of sample-multiplexed scRNA-seq, consisting of cDNA (transcriptome) and sample barcode read files: Three-sample multiplexing experiment (JS009) is a MULTI-seq dataset containing mesenchyme embryonic hind limb bud cells, embryonic stem (ES) cells, and NIH3T3 cells. Each cell sample was labelled with a distinct MULTI-seq barcode. The barcode sequences were, CATAGAGC, TCCTCGAA, and GTGTACCT for the limb bud mesenchyme cells, the ES cells, and the NIH3T3 cells, respectively. Two-sample multiplexing experiment (JS010) is a CellPlex detaset, containing ES cells and NIH3T3 cells. Each cell sample was labelled with the 3'CellPlex Kit (10X Genomics). NIH3T3 cells and ES cells were labelled with CMO301 and CMO302, respectively.

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