Takara MaxiSeq™ Human FFPE TCR+BCR (with UMIs)

MaxiSeq Human FFPE TCR+BCR (with UMIs) generates sequencing-ready libraries for both TCR and BCR sequences from 200 ng–1 µg human FFPE RNA of varying quality (RIN ≥4, DV200 ≥40%).

High-quality immune profiling libraries were prepared using MaxiSeq Human FFPE TCR+BCR (with UMIs) from highly degraded renal cell carcinoma FFPE tissue. Panel A.H&E stain of renal cell carcinoma FFPE tissue.Panel B.TCR clonotypes found in renal cell carcinoma FFPE tissue.Panel C.Two replicates show high reproducibility. RNA input was 200 ng.

MaxiSeq Human FFPE TCR+BCR (with UMIs) technology.The workflow incorporates UMIs during the reverse transcription (RT) step, enabling accurate CDR3 sequencing and minimizing PCR amplification bias. Gene-specific oligo-primed first-strand cDNA synthesis is followed by second-strand synthesis and two successive rounds of PCR amplification. The first PCR uses primers that are specific for the 5′ and 3′ universal sequences added during cDNA and second-strand synthesis. A second round of PCR was performed to add library indexes, ultimately generating a sequencing-ready library with high multiplexing capacity. After post-PCR purification, size selection, and quality analysis, the library is ready for sequencing.

MaxiSeq Human FFPE TCR+BCR (with UMIs) profiled BCR clonotypes in TILs with high reproducibility.Two replicates of two different samples of FFPE TILs were profiled. More BCR clonotypes were detected in sample 2 (Panel A), likely due to the higher lymphocyte content (Panel B). For the replicates across all samples, the correlation is >0.96 (R2values). Overlap between the replicates is low, with a Jaccard index of 0.33–0.47 (Panels C and D).