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Targeting individual cells by barcode in pooled sequence libraries

ABSTRACT: Targeting individual cells by barcode in pooled sequence libraries

PROVIDER: PRJNA479863 | ENA |

REPOSITORIES: ENA

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Targeting individual cells by barcode in pooled sequence libraries

Project description:Purpose: Selectively sequence desired single cells from pooled single cell libraries

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Targeting individual cells by barcode in pooled sequence libraries.

Project description:Transcriptional profiling of thousands of single cells in parallel by RNA-seq is now routine. However, due to reliance on pooled library preparation, targeting analysis to particular cells of interest is difficult. Here, we present a multiplexed PCR method for targeted sequencing of select cells from pooled single-cell sequence libraries. We demonstrated this molecular enrichment method on multiple cell types within pooled single-cell RNA-seq libraries produced from primary human blood cells. We show how molecular enrichment can be combined with FACS to efficiently target ultra-rare cell types, such as the recently identified AXL+SIGLEC6+ dendritic cell (AS DC) subset, in order to reduce the required sequencing effort to profile single cells by 100-fold. Our results demonstrate that DNA barcodes identifying cells within pooled sequencing libraries can be used as targets to enrich for specific molecules of interest, for example reads from a set of target cells.

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Single cell RNA-sequencing of T cells to examine barcode swapping

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Project description:HIFI-Barcode-se400

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