Targeted next-generation sequencing by specific capture of multiple genomic loci using low-volume microfluidic DNA arrays |
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Authors: | Stephan Bau Nadine Schracke Marcel Kränzle Haiguo Wu Peer F Stähler Jörg D Hoheisel Markus Beier Daniel Summerer |
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Institution: | (1) febit biomed gmbh, Im Neuenheimer Feld 519, 69120 Heidelberg, Germany;(2) febit Inc., 99 Hayden Ave, Lexington, MA 02421, USA;(3) febit holding gmbh, Im Neuenheimer Feld 519, 69120 Heidelberg, Germany;(4) Deutsches Krebsforschungszentrum, Im Neuenheimer Feld 580, 69120 Heidelberg, Germany |
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Abstract: | We report a flexible method for selective capture of sequence fragments from complex, eukaryotic genome libraries for next-generation
sequencing based on hybridization to DNA microarrays. Using microfluidic array architecture and integrated hardware, the process
is amenable to complete automation and does not introduce amplification steps into the standard library preparation workflow,
thereby avoiding bias of sequence distribution and fragment lengths. We captured a discontiguous human genomic target region
of 185 kb using a tiling design with 50mer probes. Analysis by high-throughput sequencing using an Illumina/Solexa 1G Genome
Analyzer revealed 2150-fold enrichment with mean per base coverage between 4.6 and 107.5-fold for the individual target regions.
This method represents a flexible and cost-effective approach for large-scale resequencing of complex genomes.
Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.
Stephan Bau and Nadine Schracke contributed equally to this work. |
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Keywords: | Next-generation sequencing Sequencing-by-synthesis Microarrays Microfluidics Sequence enrichment gDNA Library preparation |
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