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High-throughput automated microfluidic sample preparation for accurate microbial genomics.


ABSTRACT: Low-cost shotgun DNA sequencing is transforming the microbial sciences. Sequencing instruments are so effective that sample preparation is now the key limiting factor. Here, we introduce a microfluidic sample preparation platform that integrates the key steps in cells to sequence library sample preparation for up to 96 samples and reduces DNA input requirements 100-fold while maintaining or improving data quality. The general-purpose microarchitecture we demonstrate supports workflows with arbitrary numbers of reaction and clean-up or capture steps. By reducing the sample quantity requirements, we enabled low-input (?10,000 cells) whole-genome shotgun (WGS) sequencing of Mycobacterium tuberculosis and soil micro-colonies with superior results. We also leveraged the enhanced throughput to sequence ?400 clinical Pseudomonas aeruginosa libraries and demonstrate excellent single-nucleotide polymorphism detection performance that explained phenotypically observed antibiotic resistance. Fully-integrated lab-on-chip sample preparation overcomes technical barriers to enable broader deployment of genomics across many basic research and translational applications.

SUBMITTER: Kim S 

PROVIDER: S-EPMC5290157 | biostudies-literature | 2017 Jan

REPOSITORIES: biostudies-literature

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High-throughput automated microfluidic sample preparation for accurate microbial genomics.

Kim Soohong S   De Jonghe Joachim J   Kulesa Anthony B AB   Feldman David D   Vatanen Tommi T   Bhattacharyya Roby P RP   Berdy Brittany B   Gomez James J   Nolan Jill J   Epstein Slava S   Blainey Paul C PC  

Nature communications 20170127


Low-cost shotgun DNA sequencing is transforming the microbial sciences. Sequencing instruments are so effective that sample preparation is now the key limiting factor. Here, we introduce a microfluidic sample preparation platform that integrates the key steps in cells to sequence library sample preparation for up to 96 samples and reduces DNA input requirements 100-fold while maintaining or improving data quality. The general-purpose microarchitecture we demonstrate supports workflows with arbit  ...[more]

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