Qiagen, 334165, QIAseq Targeted cfDNA Ultra UDI Set A (96)

Box containing unique dual-indexed adapters for indexing up to 96 samples for QIAseq Targeted cfDNA Ultra Panel sequencing on Illumina platforms; 1st of 4 sets required for multiplexing up to 384 samples

Features

- Generate libraries from as little as 5 ng to 80 ng of cfDNA
- Detect variant allele frequency (VAF) down to 0.1% with >99% specificity
- Ultralow error rate with high-fidelity chemistry coupled with error-correction data analysis
- A single-day workflow with <2 hours of hands-on time
- Automation-friendly workflow

Product Details

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.

Performance

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.

Principle

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.

Procedure

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.

Applications

Applications include early cancer profiling, minimal residual disease (MRD), or research aspects of treatment resistance.