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Stephan Lorenz Wellcome Trust Sanger Institute, United Kingdom |
Abstract
The advent of single-cell sequencing technologies enabled a new era of biological research. Researchers aim to examine 1,000s of cells to understand the composition and function of cell populations and how individual cells contribute to the greater picture of tissue and organ function, development and disease. Delivering experiments on such scale is a challenging task. Manual execution of the required protocols is laborious and today’s off-the-shelf microfluidic platforms offer limited throughput. By using a combination of flow cytometry, high-speed dispensers, traditional liquid handlers and game-changing acoustic dispensing technology, we established a pipeline that delivers thousands of high-quality single-cell genomes and transcriptomes per day with a turnaround time of 2 days. Economic feasibility was achieved by minimising all assay volumes, in particular library preparation with the Nextera protocol. A conventional Nextera tagmentation and NGS library generation reaction is performed in a 50µl reaction that costs £70. By using acoustic dispensing devices, such as the ECHO 525, a scale-down by 50 – 100x can be achieved with minor modifications of the protocol, leading to significant cost reduction without sacrificing data quality. Furthermore, we demonstrate the superiority of contact-free liquid transfers in molecular biology applications with regard to reproducibility and contamination prevention. Finally, we demonstrate how acoustic dispensing can be integrated with other technologies, such as multicolour flow cytometry, to deliver next-generation experiments.
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