AmpliSeq™ for Illumina Oncology Panels on the iSeq™ 100 System
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Prep are Lib rary | Seq uence | Analyze Data AmpliSeq™ for Illumina Oncology Panels on the iSeq™ 100 System Proven amplicon chemistry meets industry-leading sequencing technology for cancer-related targeted resequencing applications. Highlights l Highly accessible sequencing solution Low cost, easy to implement, NGS system Relevant gene content for cancer research Expertly selected, cancer-focused content High analytical sensitivity with low DNA input Accurate variant detection as low as 5% allele frequency, using 1–100 ng input DNA or RNA Validated workflow for easy implementation Streamlined, comprehensive workflow with minimal hands-on time and automatic variant calling Introduction Next-generation sequencing (NGS) has transformed cancer research, enabling variant detection in numerous genes in a single assay. By focusing on genes related to a particular disease or pathway, targeted sequencing offers the ability to conduct NGS on smaller platforms, while also providing the advantage of higher depth of coverage on targeted genes. The increased analytical sensitivity that comes with higher coverage may be particularly important for detection of lowfrequency variants in heterogenous tumor tissues. The iSeq 100 System was designed to execute small-scale NGS applications. With immediate access to run various types of experiments on the benchtop, the iSeq 100 System provides autonomy and flexibility to small laboratories. The system is well suited for targeted sequencing applications that focus on diseaserelated genes. This application note demonstrates the performance of the iSeq 100 System with four distinct targeted resequencing panels, each designed with proven amplicon chemistry to focus on different aspects of cancer research: AmpliSeq for Illumina Cancer Hotspot Panel v2 is designed to detect somatic mutations across the hotspot regions of 50 genes with known associations to cancer. Starting with as little as 1 ng DNA from high-quality DNA, or DNA from formalin-fixed, paraffin-embedded (FFPE) tissues, the panel enables analytically sensitive mutation detection within multiple cancer types, including lung, colon, breast, ovarian, melanoma, and prostate. The low-input requirement enables use with precious samples. AmpliSeq for Illumina Focus Panel was developed for biomarker analysis in both DNA and RNA concurrently. The panel enables high analytical sensitivity for mutation detection in 52 genes with known relevance to solid tumors, including lung, colon, breast, ovarian, melanoma, and prostate. The Focus Panel allows quick and accurate assessment of genomic variation for translational and clinical research studies. AmpliSeq for Illumina BRCA Panel is a DNA sequencing panel that detects somatic and germline mutations across all exonic regions and the flanking intronic sequences of the BRCA1 and BRCA2 genes. BRCA1 and BRCA2 are tumor suppressor genes that have been implicated in an increased risk for breast and ovarian cancers when carrying specific mutations.1 Understanding BRCA status within the tumor may be a factor when researching potential therapies.2 Figure 1: Simple, integrated workflow—Illumina offers integrated library preparation, sequencing, and automated data analysis, creating a streamlined workflow that can go from DNA/RNA input to data in 25 hours. For Research Use Only. Not for use in diagnostic procedures.

AmpliSeq for Illumina Immune Response Panel provides detection of biomarkers associated with different leukocyte subsets, antigen presentation, checkpoint pathways, and tumor progression. Starting with 1-100 ng input RNA, the Immune Response Panel enables quick and accurate analysis of 395 genes involved in tumor-immune system interactions, and evaluation of biomarkers that can be used for developing new immunotherapy treatments. High operational efficiency The integrated workflow (Figure 1) includes AmpliSeqfor Illumina PCR-based library preparation, Illumina sequencing by synthesis (SBS)...

Prep are Lib rary | Seq uence | Analyze Data Reliable, accurate performance Learn more Combining the high coverage of targeted sequencing with the performance of the iSeq 100 System provides cancer researchers with high confidence for accurate variant detection. Using AmpliSeq chemistry to enrich for relevant targets, followed by Illumina sequencing chemistry, variant calling as low as 5% is possible with true variants distinguishable from background noise (Figure 3). To learn more about the iSeq 100 System, visit www.illumina.com/systems/sequencing-platforms/iseq.html To learn more...