QIAseq RNA Fusion XP Custom Panel (96)

QIAGEN's QIAseq RNA Fusion XP Custom Panel enables combined analysis of gene fusion detection, gene expression analysis and single nucleotide variant (SNV) calling from a single sample. The panels use single primer extension (SPE) chemistry, which requires just one gene-specific primer per target, along with unique molecular index (UMI) technology. All kits are optimized for use with unique dual index (UDI) adapters for Illumina sequencing systems. Panels are fully customizable and support both gene-level and transcript-based designs, with catalogued panels including focused targets for sarcoma, leukemia, lymphoma and other oncology applications.

In an analysis using only 5 ng of control RNA as input, the panels successfully detected eighteen clinically relevant RNA gene fusions. A built-in internal reference control enables normalization of expression levels across varying amounts of sample input. Using GM12878 RNA samples, the panels achieved 93.4% sensitivity for SNPs (VMF >=5%) within the covered and expressed target region, and can also detect SNPs in exoRNA.

The workflow begins with reverse transcription of isolated total RNA (10 ng or less) into first-strand cDNA, followed by second-strand cDNA synthesis. Adaptor complexes containing UMIs and sample indices are incorporated into the resulting dsDNA, targets of interest are enriched using SPE chemistry, and libraries are amplified via fast universal PCR with a second index added for dual indexing.