AI Article Synopsis
- Targeted genome enrichment is utilized to enhance DNA sequencing efficiency, using a new scalable protocol based on the Selector technique.
- The method improves coverage and is compatible with next-generation sequencing (NGS) for better analysis of cancer-related genes.
- Results showed 94% specificity and 98% coverage in various tissue samples, with high reproducibility and the ability to detect DNA variations, all achieved in under 24 hours without special equipment.
Article Abstract
Targeted genome enrichment is a powerful tool for making use of the massive throughput of novel DNA-sequencing instruments. We herein present a simple and scalable protocol for multiplex amplification of target regions based on the Selector technique. The updated version exhibits improved coverage and compatibility with next-generation-sequencing (NGS) library-construction procedures for shotgun sequencing with NGS platforms. To demonstrate the performance of the technique, all 501 exons from 28 genes frequently involved in cancer were enriched for and sequenced in specimens derived from cell lines and tumor biopsies. DNA from both fresh frozen and formalin-fixed paraffin-embedded biopsies were analyzed and 94% specificity and 98% coverage of the targeted region was achieved. Reproducibility between replicates was high (R(2) = 0, 98) and readily enabled detection of copy-number variations. The procedure can be carried out in <24 h and does not require any dedicated instrumentation.
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| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3025563 | PMC |
| http://dx.doi.org/10.1093/nar/gkq1005 | DOI Listing |
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