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Full-length, single cell RNA-sequencing of human bone marrow subpopulations reveals hidden complexity

ABSTRACT: Full-length, single cell RNA-sequencing of human bone marrow subpopulations reveals hidden complexity

PROVIDER: PRJNA750817 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR15346702_subreads.fastq.gz	Fastqsanger.gz
	SRR15346703_subreads.fastq.gz	Fastqsanger.gz
	SRR15304567_1.fastq.gz	Fastqsanger.gz
	SRR15304567_2.fastq.gz	Fastqsanger.gz
	SRR15304568_1.fastq.gz	Fastqsanger.gz

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Full-length, single cell RNA-sequencing of human bone marrow subpopulations reveals hidden complexity

Project description:Here, we investigate the extent to which individual cells within the total bone marrow (tot-BM) and the lineage negative (Lin-neg) populations exhibit this isoform diversity and whether distinct subpopulations in tot-BM and Lin-neg cells intersect based on transcript isoform usage. We extracted healthy donor human bone marrow tissues from discarded harvesting filters. From this total bone marrow cell preparation (tot-BM) we enriched for lineage-negative progenitor cells (Lin-neg) by magnetic selection as described earlier (Deslattes Mays et al. 2019). We then analyzed total and Lin-neg cell populations by droplet-based single cell RNA sequencing (10xGenomics). In order to increase the number of mRNA molecules detectable per cell we reduced the cell input to approximately 500 cells for each experiment. After single-cell selection, the barcoded cDNA libraries were equally divided and each pool was analyzed in parallel by short-read sequencing (Illumina) and by single-molecule real-time (SMRT) full-length RNA sequencing (Pacific Biosciences).

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