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Dual indexed library design enables compatibility of in-Drop single-cell RNA-sequencing with exAMP chemistry sequencing platforms

Austin N. Southard-Smith, Alan J. Simmons, Bob Chen, Angela L. Jones, Marisol A. Ramirez Solano, Paige N. Vega, Cherie' R. Scurrah, Yue Zhao, Michael J. Brenan, Jiekun Xuan, Martha J. Shrubsole, Ely B. Porter (+5 others)
2020 BMC Genomics  

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The increasing demand of single-cell RNA-sequencing (scRNA-seq) experiments, such as the number of experiments and cells queried per experiment, necessitates higher sequencing depth coupled to high data quality. New high-throughput sequencers, such as the Illumina NovaSeq 6000, enables this demand to be filled in a cost-effective manner. However, current scRNA-seq library designs present compatibility challenges with newer sequencing technologies, such as index-hopping, and their ability to
more » ... rate high quality data has yet to be systematically evaluated. Here, we engineered a dual-indexed library structure, called TruDrop, on top of the inDrop scRNA-seq platform to solve these compatibility challenges, such that TruDrop libraries and standard Illumina libraries can be sequenced alongside each other on the NovaSeq. On scRNA-seq libraries, we implemented a previously-documented countermeasure to the well-described problem of index-hopping, demonstrated significant improvements in base-calling accuracy on the NovaSeq, and provided an example of multiplexing twenty-four scRNA-seq libraries simultaneously. We showed favorable comparisons in transcriptional diversity of TruDrop compared with prior inDrop libraries. Our approach enables cost-effective, high throughput generation of sequencing data with high quality, which should enable more routine use of scRNA-seq technologies.
doi:10.1186/s12864-020-06843-0 pmid:32616006 pmcid:PMC7331155 fatcat:zrlmoddvcfhupi24fgt6hzdctm
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Austin N. Southard-Smith, Alan J. Simmons, Bob Chen, Angela L. Jones, Marisol A. Ramirez Solano, Paige N. Vega, Cherie' R. Scurrah, Yue Zhao, Michael J. Brenan, Jiekun Xuan, Martha J. Shrubsole, Ely B. Porter, Xi Chen, Colin J. H. Brenan, Qi Liu, Lauren N. M. Quigley, Ken S. Lau. "Dual indexed library design enables compatibility of in-Drop single-cell RNA-sequencing with exAMP chemistry sequencing platforms." BMC Genomics 21.1 (2020) 456