Abstract 5361: Predesigned gene content for rapid deployment of custom oncology panels
| Autor: | Scott V. Adams, Cynthia L. Hendrickson, Theodore B. Davis, Noa Henig, Fiona A. Stewart, Kruti M. Patel, Sarah K. Bowman, Andrew Barry, Amy B. Emerman, Eileen T. Dimalanta, Charles Elfe |
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| Rok vydání: | 2017 |
| Předmět: | |
| Zdroj: | Cancer Research. 77:5361-5361 |
| ISSN: | 1538-7445 0008-5472 |
| Popis: | Efficient utilization of targeted gene panels for oncology research is challenged by the wide variation in gene constituents specific to a given study. While focused gene panels efficiently provide the necessary depth of coverage for low frequency variant detection, the high costs and design challenges associated with de novo panel design present challenges. The NEBNext DirectTM technology utilizes a novel approach to selectively enrich nucleic acid targets ranging from a single gene to several hundred genes, without sacrificing specificity. The approach rapidly hybridizes both strands of genomic DNA with biotinyated probes prior to strepdavidin bead capture, enzymatic removal of off-target sequence, and conversion of captured molecules into sequencer-ready libraries. This results in a unique read coverage profile that results in uniform coverage across a given target. Unlike alternative hybridization methods, the approach does not necessitate upfront library preparation, and instead converts the captured molecules into dual-indexed illumina sequencer compatible libraries containing an 8 basepair sample ID and a 12bp Unique Molecule Index (UMI). The UMI individually tags each molecule prior to the final PCR amplification of the library, enabling identification of PCR duplicate molecules. The result is a 1-day protocol that enables the preparation of sequence-ready libraries from purified genomic DNA specific to the gene content included in the panel. We have designed and developed baits specific to the full exonic content of 450 genes associated with cancer. These are designed, balanced, and pooled on a per gene basis, and can be combined into customized panels, allowing rapid turnaround of specific custom gene subsets. Here, we present the ability to rapidly deploy custom gene panels across a variety of panel sizes and content, while maintaining high specificity, uniformity of coverage across target content, and sensitivity to detect nucleic acid variants from tumor samples. Citation Format: Andrew J. Barry, Amy Emerman, Sarah Bowman, Kruti Patel, Eileen Dimalanta, Scott Adams, Noa Henig, Fiona Stewart, Cynthia Hendrickson, Theodore Davis, Charles Elfe. Predesigned gene content for rapid deployment of custom oncology panels [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 5361. doi:10.1158/1538-7445.AM2017-5361 |
| Databáze: | OpenAIRE |
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