QIAseq xHYB Trinity workflow

QIAseq xHYB Trinity Kits provide hybrid capture-based target enrichment for use with Element Biosciences’ Trinity sequencing workflow on the AVITI platform.

This approach combines the modularity of the QIAseq Multimodal DNA/RNA Library Kit with our expertly curated QIAseq xHYB CGP Panels to support comprehensive genomic profiling from DNA, RNA, or both in a single research workflow.  

With Trinity technology, library prep becomes even more streamlined: A short, 30 minute hybridization step replaces the longer traditional protocol, and all manual post-hybridization steps are removed (including post-hybridization bead cleanup and amplification). This reduces hands-on time and significantly shortens the overall workflow. 

Whether you're working with limited or degraded FFPE samples, cell-free DNA or high-quality nucleic acids, this system delivers reliable and reproducible results. Using AVITI-compatible adapters and an optimized hybridization protocol, you can go from nucleic acid extraction to sequencing in 1 to 1.5 days.   

This workflow makes it easy to interrogate DNA and/or RNA alterations, including SNVs, indels, fusions and splice variants, in a single assay, dramatically reducing turnaround time and improving performance compared to a standard workflow.

• High specificity and uniformity
• Consistent target coverage across panels and sample types
• Accurate variant detection across sample types and inputs
• Sensitive detection of genomic alterations (SNVs, CNVs, fusions, exon-skipping events) and genomic signatures (TMB, MSI) using QIAseq xHYB CGP Panels for research purposes

Hybrid capture is widely used in cancer research when large genomic regions or high gene numbers need to be interrogated. However, conventional protocols are often time-consuming and involve multiple manual steps, such as bead cleanups that require strict temperature control and post-hybridization amplification. 

The QIAseq xHYB Trinity workflow simplifies library preparation from both DNA and RNA using QIAseq Multimodal DNA/RNA Library Kits and streamlines target enrichment by eliminating all manual post-hybridization steps. Libraries can be used directly with the AVITI sequencing platform without additional cleanups or amplification. 

This results in a significantly shorter workflow, so you can go from DNA to sequencer in as little as 6 hours, while maintaining high performance. The kits deliver improved on-target rates and increased recovery of original input molecules, as measured using integrated unique molecular indices (UMIs).

The workflow is streamlined for maximum efficiency and performance: 

1. Nucleic acid extraction: Use an appropriate QIAGEN extraction kit (e.g., AllPrep DNA/RNA FFPE Kit). Most QIAGEN extraction kits produce high quality nucleic acids that should be compatible with NGS. Please contact QIAGEN Technical Services for more information.
2. Library preparation: Use QIAseq Multimodal DNA/RNA Library Kits to generate high-complexity libraries from DNA and/or RNA. When building your DNA library, you can also choose between enzymatic and mechanical fragmentation.
3. Trinity hybrid capture enrichment: For comprehensive genomic profiling for your research, use QIAseq xHYB CGP Panels to enrich targets from DNA and/or RNA. 
4. Sequencing: Hybridized libraries can be loaded directly onto the AVITI using the Trinity sequencing workflow. 
5. Data analysis: Analyze data using QIAGEN Digital Insights tools for variant calling, fusion detection and annotation in your cancer research workflow. 

The QIAseq xHYB Trinity Kits, together with QIAseq Multimodal DNA/RNA Library Kits and QIAseq xHYB CGP Panels, enable comprehensive genomic profiling for cancer research: 

• Assess TMB and MSI to optimize immunotherapy outcomes
• Monitor shifts in mutations and fusions over time to study tumor heterogeneity and resistance
• Consolidate comprehensive genomic data into one assay for streamlined research efficiency
• Detect actionable alterations to validate targeted treatment efficacy