QIAseq HRR Panel

The QIAseq HRR Panel has been developed as a complete Sample to Insight solution to expand HRR biomarkers beyond BRCA1 and BRCA2. The panel's digital DNA sequencing is a unique approach to detect low-frequency variants with high confidence by overcoming the issues of PCR duplicates, false positives and library bias.

The panel is a one-box, NGS platform-agnostic solution that contains all the necessary components to construct libraries from enriched genomic targets. Primer design is based on single primer extension, in which each genomic target is enriched by one target-specific primer and a universal primer, a strategy that removes conventional two target-specific primer design restriction and reduces the amount of required primers. All primers required for a panel are pooled into an individual primer pool to reduce panel handling and number of pools required for enrichment and library construction. . The unique buffer and enzyme system used in the QIAseq HRR panel has been optimized to achieve high coverage of GC-rich genomic regions. Platform-specific indexes, which are contained in a separate box, allow the multiplexing of up to 384 samples per sequencing run.

Accuracy: Innovative digital sequencing (incorporating unique molecular indices) eliminates PCR duplication and amplification artifacts to detect low-frequency variants with high confidence.
Specificity: The unique combination of our proprietary primer design algorithm and rigorous testing of every primer assay guarantees high specificity and accurate results.
Uniformity: The QIAseq HRR Panel workflow has been optimized to deliver highly uniform sequencing results, to ensure sequencing capacity is utilized very efficiently.
Sensitivity: Digital DNA sequencing approach is optimized to deliver high confidence in calling low-frequency DNA variants.
Universality: The chemistry used in the QIAseq HRR panel and workflow is compatible with both regular and GC-rich genomic regions, allowing one to achieve 100% coverage of genes rich in GC content.
Flexibility: The QIAseq HRR panel offers a high degree of flexibility in content and sample multiplexing. Up to 384 samples can be multiplexed using the QIAseq indexes.

PCR duplicates are a major issue in targeted DNA sequencing, since, through PCR amplification, they turn unique DNA molecules into identical DNA molecules that cannot be distinguished from each other. In addition, errors from PCR amplification and sequencing process may also be present in final reads that lead to false positive variants in sequencing results. This, in turn, results in the inability to confidently call DNA variants present at low frequencies in the starting DNA material. To overcome the issue of PCR duplicates and amplification artifacts, the QIAseq HRR Panel uses digital sequencing by incorporating unique molecular indices into the starting DNA material before any amplification takes place, thereby preserving the uniqueness of the starting DNA molecules and overcoming the issues of PCR duplicates, false positives and library bias.

The entire workflow of the QIAseq HRR Panel from extracted DNA to sequencing-ready libraries can be completed in 9 hours. Extracted DNA is fragmented, genomic targets are molecularly barcoded and enriched, and libraries are constructed. Sequencing files can be fed into the QIAseq pipeline, a cloud-based data analysis pipeline, which will filter, map and align reads, as well as count unique molecular barcodes associated with targeted genomic regions, and call variants with a barcode-aware algorithm.

The QIAseq HRR Panel can be used with a wide range of sample types for numerous applications.
DNA variants:
• SNPs, InDels, and CNVs
Sample types
• FFPE
• Plasma/serum
• Fresh or frozen tissue
• Cell lines
Applications:
• Profling key 15 homologous recombination repair (HRR) genes that were part of the PROfound study