Detecting somatic genetic alterations in tumor specimens by exon capture and massively parallel sequencing Journal Article
| Authors: | Won, H. H.; Scott, S. N.; Brannon, A. R.; Shah, R. H.; Berger, M. F. |
| Article Title: | Detecting somatic genetic alterations in tumor specimens by exon capture and massively parallel sequencing |
| Abstract: | Efforts to detect and investigate key oncogenic mutations have proven valuable to facilitate the appropriate treatment for cancer patients. The establishment of high-throughput, massively parallel "next-generation" sequencing has aided the discovery of many such mutations. To enhance the clinical and translational utility of this technology, platforms must be high-throughput, cost-effective, and compatible with formalin-fixed paraffin embedded (FFPE) tissue samples that may yield small amounts of degraded or damaged DNA. Here, we describe the preparation of barcoded and multiplexed DNA libraries followed by hybridization-based capture of targeted exons for the detection of cancer-associated mutations in fresh frozen and FFPE tumors by massively parallel sequencing. This method enables the identification of sequence mutations, copy number alterations, and select structural rearrangements involving all targeted genes. Targeted exon sequencing offers the benefits of high throughput, low cost, and deep sequence coverage, thus conferring high sensitivity for detecting low frequency mutations. © JoVE 2006-2013. All Rights Reserved. |
| Keywords: | genetics; neoplasms; diagnosis; molecular biology; molecular diagnostic techniques; massively parallel sequencing; dna mutations; ffpe; high-throughput nucleotide sequencing; hybridization capture; issue 80; targeted exon sequencing |
| Journal Title: | Journal of Visualized Experiments |
| Issue: | 80 |
| ISSN: | 1940-087X |
| Publisher: | MYJoVE Corporation |
| Date Published: | 2013-10-18 |
| Start Page: | e50710 |
| Language: | English |
| DOI: | 10.3791/50710 |
| PROVIDER: | scopus |
| PUBMED: | 24192750 |
| PMCID: | PMC3947962 |
| DOI/URL: | |
| Notes: | --- - Video Article - "Export Date: 2 December 2013" - "Source: Scopus" |
Altmetric
Citation Impact
BMJ Impact Analytics
Related MSK Work