Protocol for mitochondrial variant enrichment from single-cell RNA sequencing using MAESTER.
STAR protocols
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Abstract | Single-cell RNA sequencing (scRNA-seq) enables detailed characterization of cell states but often lacks insights into tissue clonal structures. Here, we present a protocol to probe cell states and clonal information simultaneously by enriching mitochondrial DNA (mtDNA) variants from 3'-barcoded full-length cDNA. We describe steps for input library preparation, mtDNA enrichment, PCR product cleanup, and paired-end sequencing. We then detail computational steps for running maegatk, variant calling, and data integration to illuminate cell states and clonal dynamics in primary human tissues. For complete details on the use and execution of this protocol, please refer to Miller et al.. |
Year of Publication | 2025
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Journal | STAR protocols
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Volume | 6
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Issue | 1
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Pages | 103564
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Date Published | 01/2025
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ISSN | 2666-1667
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DOI | 10.1016/j.xpro.2024.103564
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PubMed ID | 39817913
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