Description
Overview
These reagents reduce background noise in sequencing data, enabling detection of low-frequency variants, such as those found in cell-free DNA (cfDNA).
Reduced error rates
Circulating tumor DNA (ctDNA) may represent a very small fraction of cfDNA, near the limit of detection for next-generation sequencing (NGS). The TruSight Oncology UMI Reagents address this challenge with UMIs and error correction software, reducing error rates to < 0.007% and enabling detection of low-frequency variants. Lower error rates increase analytical specificity, resulting in higher confidence in NGS data.
Workflow integration
The TruSight Oncology UMI Reagents include UMI adapters and indexes, plus DNA library prep and enrichment reagents. These reagents can also be paired with the TruSight Tumor 170 DNA oligos. Integration of UMIs does not create any extra steps in the library prep workflow.
Intuitive software for error correction
The UMI Error Correction App aligns reads, then uses UMIs to exclude false positives, reducing variant calling errors. The UMI Error Correction App is available in the cloud-based BaseSpace Sequence Hub or for local installation.
Example workflow
TruSight Oncology UMI Reagents
NextSeq 550 System
NovaSeq 6000 System*
BaseSpace Sequence Hub
Local UMI Error Correction App
Contact your local Illumina Support team to access the Local UMI Error Correction App.
* Performance on the NovaSeq 6000 System was demonstrated but not extensively tested.
Related applications and methods
Pathology and clinical cancer research
Our clinical cancer research solutions deliver accurate genomic information, and enable labs to analyze multiple genes in a single test.
Targeted resequencing
With targeted sequencing or resequencing, a subset of genes or a genomic region of interest is isolated and sequenced, which can conserve lab resources.
Cancer sequencing methods
NGS-based cancer sequencing methods provide more information in less time compared to single-gene and array-based approaches.
Background reduction facilitates accurate variant calling
To enable accurate detection of rare variants, UMIs are integrated with error correction software, enabling true mutations to be distinguished from background noise. The bottom panel illustrates how, by removing inherent errors that result in false positives, the reduced error rate allows true mutations (red dots) to be better distinguished from background noise (grey dots).
Reduction of error rates with UMIs
Library preparation was performed using the TruSight Oncology UMI Reagents paired with DNA content from the TruSight Tumor 170 DNA assay. 31 samples were distributed among four independent sequencing runs on the HiSeq 4000 System. Mean error rates are shown with and without collapsed reads using the UMI Error Correction App.
Sequencing Run: Mean Error Rate (Uncollapsed Reads)
1: 0.038%
2: 0.043%
3: 0.035%
4: 0.084%
TruSight Oncology UMI Reagents (16 indexes, 48 samples)
20024586
Contains the TruSight Oncology DNA library prep and enrichment reagents plus the TruSight UMI Toolkit. Provides sufficient reagents for the generation of 48 UMI-containing libraries with 16 unique index pairs. Utilized to generate libraries with reduced error rates, enabling the detection of low frequency variants. Contains no enrichment probes - TST170 content may be purchased separately. View less
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TruSight® Tumor 170 Content Set
20010188
DNA and RNA probes for 170 genes in TST170 kit. View less
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