Targeted next-generation sequencing by specific capture of multiple genomic loci using low-volume microfluidic DNA arrays
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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.
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BAU, Stephan, Nadine SCHRACKE, Marcel KRÄNZLE, Haiguo WU, Peer F. STÄHLER, Jörg D. HOHEISEL, Markus BEIER, Daniel SUMMERER, 2009. Targeted next-generation sequencing by specific capture of multiple genomic loci using low-volume microfluidic DNA arrays. In: Analytical and Bioanalytical Chemistry. 2009, 393(1), pp. 171-175. ISSN 1618-2642. eISSN 1618-2650. Available under: doi: 10.1007/s00216-008-2460-7BibTex
@article{Bau2009-01Targe-439, year={2009}, doi={10.1007/s00216-008-2460-7}, title={Targeted next-generation sequencing by specific capture of multiple genomic loci using low-volume microfluidic DNA arrays}, number={1}, volume={393}, issn={1618-2642}, journal={Analytical and Bioanalytical Chemistry}, pages={171--175}, author={Bau, Stephan and Schracke, Nadine and Kränzle, Marcel and Wu, Haiguo and Stähler, Peer F. and Hoheisel, Jörg D. and Beier, Markus and Summerer, Daniel} }
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