Automated Digital Microfluidic Sample Preparation for Next-Generation DNA Sequencing
Open Access
- 1 December 2011
- journal article
- Published by SAGE Publications in SLAS Technology
- Vol. 16 (6), 405-414
- https://doi.org/10.1016/j.jala.2011.07.001
Abstract
Next-generation sequencing (NGS) technology is a promising tool for identifying and characterizing unknown pathogens, but its usefulness in time-critical biodefense and public health applications is currently limited by the lack of fast, efficient, and reliable automated DNA sample preparation methods. To address this limitation, we are developing a digital microfluidic (DMF) platform to function as a fluid distribution hub, enabling the integration of multiple subsystem modules into an automated NGS library sample preparation system. A novel capillary interface enables highly repeatable transfer of liquid between the DMF device and the external fluidic modules, allowing both continuous-flow and droplet-based sample manipulations to be performed in one integrated system. Here, we highlight the utility of the DMF hub platform and capillary interface for automating two key operations in the NGS sample preparation workflow. Using an in-line contactless conductivity detector in conjunction with the capillary interface, we demonstrate closed-loop automated fraction collection of target analytes from a continuous-flow sample stream into droplets on the DMF device. Buffer exchange and sample cleanup, the most repeated steps in NGS library preparation, are also demonstrated on the DMF platform using a magnetic bead assay and achieving an average DNA recovery efficiency of 80% ± 4.8%.Keywords
This publication has 22 references indexed in Scilit:
- Identification of a Severe Acute Respiratory Syndrome Coronavirus-Like Virus in a Leaf-Nosed Bat in NigeriamBio, 2010
- Human Enterovirus 109: a Novel Interspecies Recombinant Enterovirus Isolated from a Case of Acute Pediatric Respiratory Illness in NicaraguaJournal of Virology, 2010
- Hydroxyapatite-Mediated Separation of Double-Stranded DNA, Single-Stranded DNA, and RNA Genomes from Natural Viral AssemblagesApplied and Environmental Microbiology, 2010
- Microarray blood testing: Pros & consBiologicals, 2010
- Sequencing technologies — the next generationNature Reviews Genetics, 2009
- A large genome center's improvements to the Illumina sequencing systemNature Methods, 2008
- Development of a digital microfluidic platform for point of care testingLab on a Chip, 2008
- Rapid droplet mixers for digital microfluidic systemsLab on a Chip, 2003
- Low voltage electrowetting-on-dielectricJournal of Applied Physics, 2002
- Electrowetting-based actuation of liquid droplets for microfluidic applicationsApplied Physics Letters, 2000