Jason Buenrostro
A Streamlined and High-Throughput Targeting Approach using Oligonucleotide-Selective Sequencing
We developed a novel approach for targeted resequencing, Oligonucleotide-Selective Sequencing (OS-Seq), in which we modify the immobilized oligonucleotide primer lawn of a next generation DNA sequencer to function as both a capture and sequencing substrate.  We designed two targeting assays in which we flanked the exons of 10 or 344 cancer genes.  In our assessment of capture performance, 87% or higher of the captured sequence originated from the target region and the target sequencing fold coverage generally fell within a 10 fold range.  To determine OS-Seq’s performance for single nucleotide variant (SNV) calling, we analyzed an individual who had undergone complete genome analysis and compared the published variants with our data with generally excellent concordance.  We also showed a similar high performance for SNV calling from the OS-Seq analysis of a matched normal colorectal cancer pair in comparison to SNP array genotyping.