Oncomine™ TCR Beta-LR Assay (Ion Torrent™)

The Oncomine TCR Beta-LR Assay is designed to measure T cell diversity and clonal expansion by sequencing T cell receptor beta chain rearrangements. This assay utilizes a newly developed long-read sequencing technology on the Ion GeneStudio S5 next-generation sequencing systems. The assay kit provides a single pool of multiplex PCR primers, library reagents, and sample barcodes. Ion AmpliSeq libraries can be generated from research samples such as RNA extracted from whole blood, fresh-frozen tissue, or FACS-sorted cells.

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The assay is designed to efficiently capture all three complementarity determining regions of the TCR beta chain (CDR1, CDR2, CDR3) with high accuracy. It enables key applications such as predictive or prognostic biomarker discovery, T cell characterization, and identification of variable gene polymorphisms. The identification of rare and abundant clones can be achieved with a flexible RNA input amount of 10 ng (minimum) and up to 1 μg. The use of RNA template allows sequencing of productive and relevant variable (V), diversity (D), and joining (J) rearrangements—improving the identification of rare clones.

Note: Information about data analysis using Ion Reporter Software v5.6 can be found in the User Guide below.

Benefits of the Oncomine TCR Beta-LR Assay include:
• Long-read RNA sequencing (up to 400 bp amplicon length) comprehensively covering TCR beta chain
• Detection of clones over a wide dynamic range
• Unbiased output generated from Ion AmpliSeq technology
• Efficient workflow with 48 hr sample-to-results time
• Flexible input requirements ranging from 10 ng to 1 μg
• Compatibility with a variety of research sample types including fresh-frozen tissue, whole blood, and sorted cells
• Streamlined and user-friendly informatics solution with multi-sample analysis functionality

Why are immunologists and immuno-oncologists interested in T cell repertoire sequencing research?
• To characterize diversity and monitor features of T lymphocytes in blood and infiltrated tumors
• To identify variable gene polymorphisms implicated in autoimmune disease or immune-mediated adverse events
• To optimize the manufacture and function of therapeutic or engineered T cells

Unlike other technologies, the Oncomine TCR Beta-LR Assay offers:
• Long-read sequencing, enabling comprehensive coverage of TCR beta chain
• High accuracy and sensitivity through a superior multiplex PCR design
• Use of very low input RNA as a starting material (10 ng)
• High multiplexing and low cost per sample, enabling large-scale studies
• Accelerated discovery cycles through fast turnaround time

The entire workflow (figure below) from isolation of RNA to analysis of samples can be accomplished in two days using the Ion Chef templating system and the Ion S5 or Ion S5 XL sequencing system.