Description
QIAseq Targeted cfDNA Ultra Kits provide a next-generation sequencing (NGS) assay for enriching selected genes and regions using 5–80 ng cell-free DNA (cfDNA). The enrichment reagent is provided as a single-tube primer mix, with up to 20,000 primers per panel. Kits are optimized for use with our QIAseq Targeted cfDNA Ultra UDI Kits for use on Illumina platforms.
Building on our expertise employing unique molecular indices (UMIs) to correct for PCR amplification bias, coupled with our proprietary highly specific target enrichment, QIAseq Targeted cfDNA Ultra Kits offer a high-fidelity library preparation for cfDNA samples that enables reliable detection of somatic variants at 0.1% VAF. When paired with QIAGEN CLC Genomics Workbench for variant calling, QIAseq Targeted cfDNA Ultra Kits can deliver high sensitivity in detecting ultralow variants with >99% specificity.
QIAseq Targeted cfDNA Ultra Kits integrate unique molecular indices (UMIs) and high-fidelity chemistry to identify rare variants with high confidence. The chemistry is further optimized with a specially formulated enrichment chemistry to achieve highly efficient enrichment of both regular and GC-rich regions at high multiplex levels (up to 20,000 primers per panel). When coupled with QIAGEN CLC Genomics Workbench for secondary analysis, QIAseq Targeted cfDNA Ultra Kits enable ultrasensitive variant detection down to 0.1% with >99% specificity. The required amount of template for a single QIAseq Targeted cfDNA Ultra sequencing reaction ranges from 5–80 ng cfDNA.
End preparation and UMI assignment First, cfDNA samples are end repaired and A-tailed within a single, controlled multienzyme reaction. The prepared DNA fragments are then ligated with a sequencing platform-specific adapter containing 12 bp UMIs. Target enrichment and final library construction For enrichment, ligated DNA molecules are subject to several cycles of targeted PCR using one region-specific primer and one universal primer complementary to the adapter. A universal PCR is ultimately carried out to amplify the library and add platform-specific adapter sequences and sample indices.
