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dCITI-Seq: droplet combinational indexed transposon insertion sequencing

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Abstract

The rapid development of high-throughput parallel sequencing poses new challenges for large-scale barcoding and sequencing library construction. Here, we present droplet combinational indexed transposon insertion sequencing (dCITI-Seq), in which samples are indexed by the direct insertion of index-containing adaptors through transposition. The random combination of two sets of adaptors with known barcodes and massively parallel transposition was realized via a robust droplet pairing and merging platform. This strategy potentially enlarges the indexing capacity and decreases index crosstalk. Also, dCITI-Seq exhibited a lower GC base preference than conventional in-tube transposition library preparation. With a custom bioinformatic processing, it could be further applied to large-scale single-cell sequencing.

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Funding

This work was supported by the Natural Science Foundation of Jiangsu Province (BK20211513), National Natural Science Foundation of China (61971125), and Six Talent Peaks Project of Jiangsu Province (2019-SWYY-004).

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Authors and Affiliations

  1. State Key Laboratory of Bioelectronics, School of Biological Science and Medical Engineering, Southeast University, Nanjing, 210096, China

    Jing Tu, Yi Qiao, Zheyun Xu, Na Lu, Naiyun Long & Zuhong Lu

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  1. Jing Tu
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  2. Yi Qiao
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  3. Zheyun Xu
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  4. Na Lu
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  5. Naiyun Long
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  6. Zuhong Lu
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Contributions

Conceptualization: Jing Tu. Investigation and methodology: Yi Qiao, Zheyun Xu, Naiyun Long. Data curation and formal analysis: Zheyun Xu, Na Lu. Resources and supervision: Jing Tu, Zuhong Lu. Writing—original draft: Jing Tu, Yi Qiao, Zheyun Xu. Writing, review, and editing: Jing Tu, Yi Qiao.

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Correspondence to Jing Tu or Zuhong Lu.

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Tu, J., Qiao, Y., Xu, Z. et al. dCITI-Seq: droplet combinational indexed transposon insertion sequencing. Anal Bioanal Chem 414, 2661–2670 (2022). https://doi.org/10.1007/s00216-022-03902-1

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  • DOI: https://doi.org/10.1007/s00216-022-03902-1

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