Highly multiplexed targeted DNA sequencing from single nuclei

Marco L. Leung, Yong Wang, Charissa Kim, Ruli Gao, Jerry Jiang, Emi Sei, Nicholas E. Navin

Research output: Contribution to journalArticlepeer-review

41 Scopus citations


Single-cell DNA sequencing methods are challenged by poor physical coverage, high technical error rates and low throughput. To address these issues, we developed a single-cell DNA sequencing protocol that combines flow-sorting of single nuclei, time-limited multiple-displacement amplification (MDA), low-input library preparation, DNA barcoding, targeted capture and next-generation sequencing (NGS). This approach represents a major improvement over our previous single nucleus sequencing (SNS) Nature Protocols paper in terms of generating higher-coverage data (>90%), thereby enabling the detection of genome-wide variants in single mammalian cells at base-pair resolution. Furthermore, by pooling 48-96 single-cell libraries together for targeted capture, this approach can be used to sequence many single-cell libraries in parallel in a single reaction. This protocol greatly reduces the cost of single-cell DNA sequencing, and it can be completed in 5-6 d by advanced users. This single-cell DNA sequencing protocol has broad applications for studying rare cells and complex populations in diverse fields of biological research and medicine.

Original languageEnglish (US)
Pages (from-to)214-235
Number of pages22
JournalNature Protocols
Issue number2
StatePublished - Feb 1 2016

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)


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