QIAseq targeted DNA Extended panel (96)

From QIAGEN, the QIAseq Targeted DNA Extended Panel is a complete Sample to Insight solution for ultrasensitive targeted next-generation sequencing (NGS) of DNA on Illumina NGS systems. The panel enables digital DNA sequencing by incorporating molecular barcodes into the starting DNA material before any amplification takes place, preserving the uniqueness of starting DNA molecules and overcoming issues of PCR duplicates, false positives, and library bias to allow confident detection of low-frequency variants.

Each panel is a one-box, NGS platform-agnostic solution containing all components needed to construct libraries from enriched genomic targets. Primer design is based on QIAseq Enrichment Technology, in which each genomic target is enriched by one target-specific primer paired with one universal primer, removing conventional two target-specific primer design restrictions and reducing the amount of required primers. All primers required for a panel are pooled into a single primer pool to reduce panel handling. Extended panels allow the content of any cataloged panel to be extended by adding new primers, which are added to the content of the cataloged panel. The unique buffer and enzyme system has been optimized to achieve high coverage of GC-rich genomic regions.

The workflow from extracted DNA to sequencing-ready libraries can be completed in 9 hours, with extracted DNA enzymatically fragmented, genomic targets molecularly barcoded and enriched, and libraries constructed. Required template ranges from 10 to 80 ng for fresh DNA or 100 to 250 ng for formalin-fixed paraffin-embedded (FFPE) DNA, with isolated DNA as low as 20 ng supported. Library fragments serve as templates for target enrichment using single primer extension, with targets enriched using a single gene-specific primer and a universal forward primer, followed by library amplification and sample indexing using the IL-S5 sample index primer and a universal primer. The panels use integrated unique molecular indices (UMIs) to facilitate ultrasensitive variant detection from cells, tissue, and biofluids, supporting identification of somatic mutations, single nucleotide polymorphisms, copy number variation, and small insertions/deletions. Sequencing files can be analyzed using the cloud-based QIAseq pipeline, which filters, maps, and aligns reads, counts unique molecular barcodes, and calls variants with a barcode-aware algorithm; data can then be fed into IVA or QCI for interpretation. The panels are intended for molecular biology applications and are not intended for the diagnosis, prevention, or treatment of a disease.