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Targeting individual cells by barcode in pooled sequence libraries.
- Source :
-
Nucleic acids research [Nucleic Acids Res] 2019 Jan 10; Vol. 47 (1), pp. e4. - Publication Year :
- 2019
-
Abstract
- 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.
Details
- Language :
- English
- ISSN :
- 1362-4962
- Volume :
- 47
- Issue :
- 1
- Database :
- MEDLINE
- Journal :
- Nucleic acids research
- Publication Type :
- Academic Journal
- Accession number :
- 30256981
- Full Text :
- https://doi.org/10.1093/nar/gky856