Principle

DNA sequencing is a useful tool to detect genetic variations, including somatic mutations, SNPs, and small insertions and deletions. 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. GeneRead QIAact Panels, Powered by QCI use multiplex PCR-based targeted enrichment technology in combination with a sophisticated primer design algorithm. This technique enables amplification and enrichment of any gene or targeted region in the human genome for detection of genetic variation using NGS. Adjacent and potentially interacting primer pairs are separated into different pools for optimal performance. GeneRead QIAact Panels, Powered by QCI are designed to analyze a panel of genes and can be used with the QIAGEN GeneReader instrument. The targeted enrichment process is essential for the efficient utilization of medium-throughput sequencers such as the QIAGEN GeneReader.
GeneRead QIAact Panels, Powered by QCI have been optimized in combination with the GeneRead DNAseq Panel PCR Kit V2 to provide superior sensitivity and linear multiplex amplification. The simplicity of the PCR method makes these panels accessible for routine use in every clinical research laboratory.
The Actionable Insights Tumor Panel is one of the GeneRead QIAact Panels, Powered by QCI and was developed with an unprecedented process. It leverages the expertly curated QIAGEN Knowledge Base, which focuses only on clinically relevant findings such as approved therapeutics labels, professional association practice guidelines and active late-stage clinical trials. The resulting design is a unique set of genes and variants with an unparalleled level of direct relevance that provides necessary and sufficient insights.

Procedure

The Actionable Insights Tumor Panel covers specific hotspots designed to detect SNVs and small (<20 bp) indels, so one 4-pool set is provided. GeneRead QIAact Panels, Powered by QCI can enrich selected genes and/or regions using as little as 40 ng genomic DNA in 3 hours for a 4-pool panel. Genomic DNA samples are combined with the primer mix and PCR reagent, and PCR is performed in a standard thermocycler. After the reaction is complete, the reactions for each sample are pooled and the enriched DNA is purified. The purified DNA is then ready for NGS library construction and sequencing using the QIAGEN GeneReader.

Applications

For targeted enrichment prior to next-generation sequencing (NGS) applications that use the QIAGEN GeneReader® instrument