GeneRead QIAact AIT DNA UMI Panel
For even more actionable solid tumor insights from FFPE samples
The GeneRead QIAact AIT DNA UMI Kit builds on the success of the GeneRead QIAact Actionable Insights Tumor Panel. Covering the most cancer relevant variants in 30 genes implicated in the cancer types that can most benefit from genetic testing, this panel is designed to deliver more actionable cancer research insights than ever before. Incorporating unique molecular index (UMI) technology, you can now detect not just SNVs and Indels, but also CNVs with uniform coverage and sensitive variant detection guaranteed.
Integrated as part of the complete GeneReader NG System workflow, the panel offers a complete assay, including QCI Analyze and Interpret for seamless variant reporting based on ACMG and the new AMP/ASCO/CAP guidelines for somatic variant classification.
The GeneRead QIAact AIT DNA UMI Panel is for Research Use Only. Not for use in diagnostic procedures.
In performance tests with FFPE samples on the GeneReader NGS System, the GeneRead QIAact Actionable Insights Tumor Panel showed excellent coverage at all variant positions, even with FFPE samples of up to 20 years old. In a comparative analysis of RAS variant detection from FFPE samples, results generated with this panel on the GeneReader NGS System were 100% concordant with those obtained using QIAGEN therascreen PCR assay and pyrosequencing. Additionally, these data are also entirely consistent with those obtained on an Illumina MiSeq sequencer.
DNA sequencing is a useful tool to detect genetic variations, including somatic mutations, single nucleotide variants (SNVs), copy number variation (CNVs) and small insertions and deletions (inDels). Targeted enrichment technology enables next-generation sequencing (NGS) platform users to sequence specific regions of interest instead of the entire genome, effectively increasing sequencing depth and throughput with lower cost. Existing target enrichment methods, library preparation and sequencing steps all utilize enzymes and amplification processes, which introduce substantial bias and artifacts. These biases and artifacts lead to background artefactual errors that greatly limit the detection of true low-frequency variants in heterogeneous samples such as tumors.
The GeneRead QIAact AIT DNA UMI Kit integrates unique molecular index (UMI) technology into a gene-specific, primer-based target enrichment process, enabling sensitive variant detection of targeted genomic regions by NGS on the GeneReader system.
The GeneRead QIAact AIT DNA UMI Kit has been optimized in combination with a specially formulated enrichment chemistry to achieve highly efficient enrichment on both regular and GC-rich regions at high multiplex levels.
The GeneRead QIAact AIT DNA UMI Kit is provided as two primer mix tubes, with approximately 500 primers per tube. The kit is designed to enrich specific target regions in select genes (AKT1, ALK1, BRAF, CTNNB1, DDR2, EGFR, ERBB2, ERBB3, ERBB4, ESR1, FBXW7, FGFR1, FGFR2, FGFR3, FLT3, GNA11, GNAQ, HRAS, KIT, KRAS, MAP2K1, MAP2K2, MET, NOTCH1, NRAS, PDGFRA, PIK3CA, RAF1, SMAD4, STK11) using 40-160 ng of DNA.
Genomic DNA samples are first fragmented, end-repaired and A-tailed using a single, controlled multi-enzyme reaction. The prepared DNA fragments are then ligated at their 5’ ends to a GeneReader specific adapter containing a UMI and a 9 base-pair (bp) sample-specific bar code.
Ligated DNA molecules are subject to limited cycles of target enrichment PCR, with gene-specific primers targeting hot spot regions paired with universal primers complimentary to an adapter sequence. Gene specific primers are tiled across the whole exon.This reaction ensures that intended targets and UMIs are enriched sufficiently to be represented in the final library. A universal PCR with GeneReader specific sequences is then carried out to amplify the targets and complete the library.
Once the library is sequenced, results can be analyzed using the GeneRead QIAact AIT DNA UMI Kit workflow, which will automatically perform all steps necessary to generate a DNA sequence variant report from your raw NGS data. All detected variants can be further interpreted by QIAGEN Clinical Insight (QCI) analysis.
For targeted enrichment prior to next-generation sequencing (NGS) applications that use the QIAGEN GeneReader instrument
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