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PLoS One. 2013 Jul 22;8(7):e68988. doi: 10.1371/journal.pone.0068988. Print 2013.

A microfluidic DNA library preparation platform for next-generation sequencing.

Author information

1
Department of Biotechnology and Bioengineering, Sandia National Laboratories, Livermore, California, USA.

Abstract

Next-generation sequencing (NGS) is emerging as a powerful tool for elucidating genetic information for a wide range of applications. Unfortunately, the surging popularity of NGS has not yet been accompanied by an improvement in automated techniques for preparing formatted sequencing libraries. To address this challenge, we have developed a prototype microfluidic system for preparing sequencer-ready DNA libraries for analysis by Illumina sequencing. Our system combines droplet-based digital microfluidic (DMF) sample handling with peripheral modules to create a fully-integrated, sample-in library-out platform. In this report, we use our automated system to prepare NGS libraries from samples of human and bacterial genomic DNA. E. coli libraries prepared on-device from 5 ng of total DNA yielded excellent sequence coverage over the entire bacterial genome, with >99% alignment to the reference genome, even genome coverage, and good quality scores. Furthermore, we produced a de novo assembly on a previously unsequenced multi-drug resistant Klebsiella pneumoniae strain BAA-2146 (KpnNDM). The new method described here is fast, robust, scalable, and automated. Our device for library preparation will assist in the integration of NGS technology into a wide variety of laboratories, including small research laboratories and clinical laboratories.

PMID:
23894387
PMCID:
PMC3718812
DOI:
10.1371/journal.pone.0068988
[Indexed for MEDLINE]
Free PMC Article

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