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AGBT19 Poster - Utilizing a novel microfluidic technology to enable robust and rapid “hands-off” library preparation for human whole genomes

Maura Costello1, Tim Desmet1, Julia Yoo2, Severine Margeridon2, Nikolay Sergeev2, Yu-Hung Chen2, Mais Jebrail2, Fay Christodoulou2

1 Broad Institute Genomics Platform, Cambridge, MA 02141

2 Miroculus, Inc., San Francisco, CA 94107

As whole genome sequencing has become more widely adopted due to ever decreasing sequencing costs, Broad Genomics has seen an increased demand for our PCR-Free genome offerings in both the traditional and clinical research spaces. For clinical research in particular, rapid turn around, complete chain of custody tracking, reproducibility, and tolerance for lower input and lower quality samples are all highly desirable features for a whole genome workflow. To address these needs, we are collaborating with Microculus to develop sample preparation methods using their novel aeros™ microfluidic technology. Our first application will be PCR-Free whole genomes utilizing a version of the KAPA Hyper Prep workflow. All sample manipulation steps including enzyme additions, incubations, and magnetic bead-based clean ups are done on the Miroculus instrument, with the output being a sequencer-ready PCR-Free library. The technology allows for hands-off library preparation, reducing the chance for user error, cross contamination, and sample swaps. Here, we demonstrate that libraries made using the Miroculus instrument are equivalent or better in terms of turnaround time, reproducibility, yield, and data quality than current manual or plate-based automated methods. Additionally, the aeros™ microfluidic technology improves the stoichiometry of library preparation reactions by introducing active mixing steps during incubations and shrinking reaction volumes, leading to higher adapter ligation conversion rates. We demonstrate this increased efficiency allows creation of robust PCR-Free libraries from a variety of sample types with much less starting DNA input than our current plate based automated methods, enabling PCR-Free genomes from samples that previously did not meet our specifications for DNA quantity. Going forward, we believe this technology has the potential to be applied to a wider variety applications, including DNA extractions, blood biopsy targeted methods, and other workflows, and could enable ultra fast rapid turn around applications for clinical research.

 

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