QIAseq Targeted RNA Panel Human TCR (12)

QIAGEN's targeted RNA-seq panel enables characterization of T-cell receptor (TCR) alpha, beta, gamma and delta genes from human or mouse samples. Designed for RNA-seq library construction, it accepts total RNA, FFPE/fragmented RNA, or enriched mRNA as input, with a recommended range of 200 pg to 1000 ng of RNA. Gene targets include human TRA, TRB, TRD and TRG, and mouse Tcra, Tcrb, Tcrd and Tcrg.

The kit incorporates unique molecular indices (UMIs) through QIAGEN Enrichment Technology, which reduces amplification bias compared to multiplexed PCR-based V and J primer pools. Before amplification, each original cDNA molecule receives a UMI containing up to 18 bases via random sequence adapter ligation, providing more than 268 million possible indices per adapter. The workflow begins with gene-specific reverse transcription (5 µL of RNA in a 6 µL cDNA reaction volume, minimum 40 pg/µL concentration), followed by second-strand synthesis, end repair with A-addition in a single-tube protocol, UMI assignment, TCR-specific enrichment, and a final universal PCR that introduces unique dual indices. Acceptable RIN values range from 1 to 10, with a minimum DV600 of 30%.

The panel is compatible with Illumina NGS systems including MiSeq, NextSeq 1000/2000 and NovaSeq 6000, with best sequencing results on 500 or 600 cycle flow cells using paired-end, 600-cycle reads on NextSeq 1000/2000. Online data analysis is included via GeneGlobe Analyze, and the QIAseq Immune Repertoire pipeline handles read mapping and clonotype calling. Required QIAseq TCR UDI index kits and the recommended QIAGEN XpressRef Universal Total RNA control are sold separately.