b'Next-Generation Sequencing (NGS) | Research Use Only (RUO) AssaysTRB Assay Next-Generation Sequencing (NGS) | RUO AssaysLymphoTrack TRB AssayAssay Uses Analysis of the rearranged TRB locus increases the probability of identifying T-cell receptor gene rearrangements, as compared to This research use only (RUO) assay for next-generation sequencingtesting for TRG gene rearrangements only. As a result, combining (NGS) identifies clonal TRB V-(D-)J rearrangements, the associatedthe analysis of TRB and TRG loci increases the sensitivity of clonality V-(D-)J region DNA sequences, it provides the frequency distributiondetection. of V, D, and J region segment utilization using the Illumina MiSeq platform. The human T-cell receptor beta (TRB) gene locus on chromosome 7 (7q34) includes 65 V (variable) gene segments, followed by two Summary and Explanation of the Test separate clusters of genes each containing a D (diversity) gene, several J (joining) genes, and a C (constant) region spread over In contrast to the conventional fragment analysis assays for clonality685 kilobases. The two C genes, TRBC1 and TRBC2, encode highly that utilize three master mixes, this NGS-based assay contains a singlehomologous products with no functional difference.multiplex master mix to target conserved V, D, and J regions of theTRB gene locus described in lymphoid malignancies. This reducesDuring lymphoid cell development, antigen receptor genes undergo DNA sample requirements and simplifies the testing workflow. somatic gene rearrangements. Specifically, during T-cell development, The LymphoTrack TRB master mix primers are also designed withgenes encoding TRB molecules are assembled from multiple Illumina adapters and 8 indices (Kit A) or 24 indices (Panel). polymorphic gene segments that generate V-D-J combinations This allows amplicons generated from different indexed TRB unique in both length and sequence.master mixes to be pooled into a single sequencing library. Since leukemias and lymphomas originate from the malignant The associated LymphoTrack Software is capable of sorting transformation of individual lymphoid cells, all leukemias and complex NGS data by gene target. This offers a second layerlymphomas generally share one or more cell-specific or clonal of multiplexing to reduce per sample testing costs by allowingantigen receptor gene rearrangements. Therefore, tests that detect amplicons from any LymphoTrack Assay (e.g. IGH, IGK, TRB, TRG) toTRB clonal rearrangements can be useful in the study of B- andbe sequenced on the same flow cell. In addition, the LymphoTrackT-cell malignancies.Software provides easy visualization of data and the LymphoTrack Minimal Residual Disease Data Analysis Tool allows identified sequences to be tracked and monitored in subsequent samples.Specimen RequirementPositive clonal and negative polyclonal DNA controls are included in50 ng of high-quality genomic DNA.kits. Please see the LymphoTrack MRD software section to learn how the LymphoTrack Assays can be applied to MRD studies, or emailmarketing@invivoscribe.com. References1.EP Rock et al., J Exp Med 179 (1): 323-8 (1994).Background2.JE Miller et al., Molecular Genetic Pathology (2nd Ed., sections The LymphoTrack TRB Assay represent a significant improvement over30.2.7.13 and 30.2.7.18). New York, USA: Springer Science &clonality assessment by fragment analysis by providing two importantBusiness Media (2013).and complementary uses:1.Detection of initial clonal populations.2.dentification of sequence information required to track clonal Irearrangements in subsequent samples.Simplified Representation of the TRB GeneV D1 J1 C1 D2 J2 C2Depicted are the variable (V), diversity (D), and joining (J) gene regions involved in TRB gene rearrangements, in addition to the downstream consensus (C) gene regions. 46'