Journal
ANALYTICA CHIMICA ACTA
Volume 1182, Issue -, Pages -Publisher
ELSEVIER
DOI: 10.1016/j.aca.2021.338954
Keywords
Next generation sequencing; Library preparation; Centrifugal microfluidics; Point-of-care; Automation
Categories
Funding
- Ministry of Economic Affairs, Labour and Housing of the State of Baden-Wurttemberg, Germany, within the project PRIMO
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Next generation sequencing is transitioning from a research tool to a method used in diagnostic routine. The complete sequencing workflow includes sample pre-processing, library preparation, sequencing, and bioinformatics analysis. This study presents an automated library preparation method for whole genome sequencing using centrifugal microfluidics, which significantly improves efficiency.
Next generation sequencing is evolving from a research tool into a method applied in diagnostic routine. The complete sequencing workflow includes sample pre-processing, library preparation, sequencing and bioinformatics. High quality in each of these steps is necessary to obtain excellent sequencing results. The tedious and error-prone library preparation poses a significant challenge for smaller laboratories, where high throughput pipetting robots are not cost-effective. Here we present an automated library preparation for whole genome sequencing using centrifugal microfluidics. Two samples can be run per cartridge. Precise metering of reagents allows the required liquid volumes to be reduced by 40% and the amount of sample used by 60%. The functionality of the cartridge is demonstrated with bacteria and DNA extracted from a human FFPE sample. For the bacterial sample, mean sequencing depths from 140 to 183 reads and a coverage of 99.8% of the reference genome were detected. For the human DNA, mean sequencing depths of 4.4-5.7 reads and a coverage of 78.2% of the effective reference genome were observed. (C) 2021 Elsevier B.V. All rights reserved.
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