GeneRead QIAact Custom Panels
For targeted sequencing of the variants most relevant to your research
Whatever your area of medical research, you can use the GeneReader NGS System to gain actionable insights with the GeneRead QIAact Custom Panels. Consult with us about your specific requirements and let us help you to develop a panel that will target the genes or regions of most value to your research†.
The GeneRead QIAact Custom Panels can be designed to detect all genetic variants, including somatic mutations, single nucleotide variants (SNVs), copy number variation (CNVs), small insertions, deletions (inDels) and fusions*. Incorporating our novel unique molecular index (UMI) technology guarantees reduced variant calling errors and provides highly sensitive variant detection. As an integral part of our Sample to Insight NGS solution, the GeneRead QIAact DNA Custom Panels are supported by QCI Analyze and Interpret, allowing seamless sequence data analysis and interpretation at the push of a button.
The GeneRead QIAact DNA Custom kit contains all of the reagents required for both the target enrichment and library preparation steps of the GeneReader NGS System workflow, to help streamline your laboratory operation.
†Dependent on feasibly assessment and capacity
The GeneRead QIAact DNA Custom Panel Kit and GeneRead QIAact Panel Cleanup Kit are for Research Use Only. Not for use in diagnostic procedures.
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 and effectively increase throughput with lower cost. The GeneRead QIAact DNA Custom panel is based on customer requirements enabling you to specify the regions of specific interest to your research (SNVs, InDels, CNVs and fusions*).
Existing target enrichment methods, library preparation and sequencing steps all utilize DNA polymerase and amplification processes, which can introduce artifacts and substantial bias. These artifacts and biases lead to background artefactual errors that greatly limit the detection of true low-frequency variants in heterogeneous samples such as tumors. The GeneRead QIAact DNA Custom Panel integrates unique molecular index (UMI) technology and a gene-specific, primer-based target enrichment process to enable sensitive variant detection of targeted genomic regions by NGS on the GeneReader system. The use of UMI technology results in higher confidence variant calling, which is reflected in the high Q scores obtainable.
The GeneRead QIAact DNA Custom Panel is provided as two primer mix tubes, with a customer specific number of primers per tube and is designed to enrich selected genes and regions using 40 ng of starting DNA.
Genomic DNA samples are first fragmented, end-repaired and A-tailed within a single, controlled multi-enzyme reaction. The prepared DNA fragments are then ligated at their 5’ ends with a specific adapter containing a UMI and a sample-specific bar code.
Ligated DNA molecules are subject to limited cycles of target enrichment PCR, with gene-specific primers targeting genomic regions and a universal primer complimentary to an adapter sequence. This reaction ensures that intended targets and UMIs are enriched sufficiently to be represented in the final library. A universal PCR is then carried out to amplify the library and add GeneReader specific sequences which completes the library.
Three DNA cleanup steps are required during this protocol for the removal of buffer and enzymes that might inhibit downstream library processing:
• A first cleanup procedure takes place after the adapter ligation
• A second cleanup procedure takes place after target enrichment PCR
• A final cleanup procedure is performed after the universal PCR step
Each of these cleanup steps is fully automatable on the QIAcube platform with the GeneRead QIAact Cleanup Kit (catalogue no. 185446) to reduce sample processing time and risk of handling errors.
Once the library is sequenced, results can be analyzed using the GeneRead QIAact DNA Custom Panel 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 constructing targeted, molecularly bar-coded libraries from DNA for digital sequencing as part of the GeneReader NGS System workflow.
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