Description
Overview
TruSight Tumor 170 is an NGS assay that assesses 170 genes associated with common solid tumors.
Assess fusions, splice variants, insertions/deletions (indels) and single nucleotide variants (SNVs), and amplifications in one assay using DNA and RNA
Detect variants at 5% allele frequency with as little as 40 ng DNA and RNA input
Maximize results with reliable small variant detection from high- and low-quality FFPE samples
Comprehensive coverage of cancer-related variants
Maximize single-assay efficiency using DNA and RNA from a single sample to assess multiple variant types, preserving valuable tissue, time, and resources. Expert-selected evidence-based content provides researchers with comprehensive coverage of variants most likely to play a role in tumorigenesis.
Combined workflow for DNA and RNA
DNA and RNA libraries are prepared, sequenced, and analyzed simultaneously for efficient assessment of numerous types of somatic variants.
Streamlined data analysis
Use the TruSight Tumor 170 App on BaseSpace Sequence Hub for sequencing data analysis and management. Local secondary analysis is available on a Docker-based software.
Applications
TruSight Tumor 170 provides efficient comprehensive coverage of cancer-related variants (small variants, amplifications, splice variants, and fusions) in RNA and DNA obtained from a single FFPE sample.
Example workflow
NextSeq 550 System
BaseSpace Sequence Hub App for TruSight Tumor 170
TruSight Tumor 170 Local App: Docker-based Software
Project recommendations
Instrument: Recommended number of samples
NextSeq 550 System: Samples per run (high output): 16 (8 DNA + 8 RNA), 10 if DNA only, 16 if RNA only
Samples per run (high output): 16 (8 DNA + 8 RNA), 10 if DNA only, 16 if RNA only
2 × 101 bp (max recommended)
Related applications and methods
Cancer panels
Access a broad range of fixed and custom next-generation sequencing panels targeting known cancer-related gene variants.
Solid tumor profiling
Next-generation sequencing (NGS) provides a comprehensive method for assessing the majority of genes associated with solid tumors.
NGS for drug development
Figures
TruSight tumor 170 targets all coding exons per the current RefSeq database1 in 170 genes. The content includes 55 genes for fusions and splice variants, 148 SNVs and indels, and 59 amplifications.
DNA and RNA samples follow the same workflow, after the cDNA synthesis step (for RNA) and the shearing step (for DNA).
DNA from FFPE samples of varying quality was extracted and evaluated using TruSight Tumor 170 and sequenced on the NextSeq 500 System. Sample quality was assessed using qPCR to measure DNA amplification potential. The ΔCq value indicates the cycle threshold (Ct) value of each DNA sample minus the Ct value of a DNA standard.
Removing cancer’s veil: TruSight Tumor 170 gene panel
Learn how a single assay interrogates both DNA and RNA to maximize the output from a single FFPE sample.
TruSight Tumor 170 Kit (24 Samples)
OP-101-1004
Includes library prep and enrichment reagents to allow for solid tumor profiling of 170 genes from FFPE samples. View less
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TruSight Tumor 170 Kit, With NextSeq v2.5 Reagents (24 Samples)
20028821
Includes library prep, enrichment, and sequencing consumables to allow for solid tumor profiling of 170 genes from FFPE samples. View less
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How does TruSight Tumor 170 use both DNA and RNA in one assay?
During library prep and enrichment, the DNA and RNA samples go through the same workflow following sample shearing (DNA) and cDNA synthesis (RNA). This results in the ability to process the DNA and RNA samples in parallel. The DNA and RNA libraries can then be sequenced on the same sequencing run.
How can I extract DNA and RNA from my FFPE tissue?
Extraction reagents are not included in the TruSight Tumor 170 kit. The QIAGEN AllPrep DNA/RNA FFPE Kit (Catalog no. 80234) has demonstrated high yields of nucleic acids in comparison to other extraction methods for this assay. The QIAGEN AllPrep DNA/RNA FFPE Kit includes a DNase I digestion step during RNA extraction. You can select other commercially available extraction kits at your discretion.
Which Illumina sequencing systems are compatible with TruSight Tumor 170?
TruSight Tumor 170 performance has been verified on the NextSeq 500 and NextSeq 550 Systems, and on the HiSeq 2500 System in rapid run mode.
How much input DNA and RNA is needed for TruSight Tumor 170?
Illumina recommends adding a minimum of 40 ng of DNA or RNA. For samples that are highly degraded, assay performance may be improved by using a maximum of 120 ng of DNA and 85 ng of RNA.
What coverage level is expected?
Coverage may vary based on sample quality and multiplexing level. Please review the DNA_SampleMetricsReport.txt and RNA_SampleMetricsReport.txt files produced by the TruSight Tumor 170 app for each analysis. This report contains a metric indicating the percent bases with > 100× coverage. A sequencing run with 8 high-quality DNA and 8 high-quality RNA samples (16 libraries) has demonstrated ≥ 99% bases with 100× coverage.
How do I analyze data?
Use the TruSight Tumor 170 app available in BaseSpace Sequence Hub cloud environment or a local Docker App to analyze TruSight Tumor 170 libraries.
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References :
1. O'Leary NA, Wright MW, Brister JR, et al. Reference sequence (RefSeq) database at NCBI: current status, taxonomic expansion, and functional annotation . Nucleic Acids Res. 2016;44(D1):D733-45.
doi:10.1093/nar/gkv1189
