期刊
GENOME BIOLOGY
卷 23, 期 1, 页码 -出版社
BMC
DOI: 10.1186/s13059-022-02615-z
关键词
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资金
- National Human Genome Research Institute/National Institute of Health Training Grant [1T32HG008345-01]
- Hellman Foundation
- Santa Cruz Cancer Benefit Group
- National Institute of General Medical Sciences/National Institute of Health Grant [R35GM133569]
In this study, we utilized R2C2 sequencing technology to analyze 12 million full-length cDNA molecules generated by the 10X Genomics platform. Independent of Illumina data, we successfully identified and generated clusters for B cells, T cells, and monocytes, as well as isoform-level transcriptomes for cells and cell types. Additionally, we extracted paired adaptive immune receptor sequences unique to each T and B cell.
High-throughput single-cell analysis today is facilitated by protocols like the 10X Genomics platform or Drop-Seq which generate cDNA pools in which the origin of a transcript is encoded at its 5' or 3' end. Here, we used R2C2 to sequence and demultiplex 12 million full-length cDNA molecules generated by the 10X Genomics platform from similar to 3000 peripheral blood mononuclear cells. We use these reads, independent from Illumina data, to identify B cell, T cell, and monocyte clusters and generate isoform-level transcriptomes for cells and cell types. Finally, we extract paired adaptive immune receptor sequences unique to each T and B cell.
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