b'Next-Generation Sequencing (NGS) | Research Use Only (RUO) AssaysTRB Assay Next-Generation Sequencing (NGS) | RUO AssaysLymphoTrack TRB AssayAssay Uses Background This research use only (RUO) assay for next-generation sequencingThe LymphoTrack TRB Assay represent a significant improvement over (NGS) identifies clonal TRB V-(D-)J rearrangements, the associatedclonality assessment by fragment analysis by providing two important V-(D-)J region DNA sequences, and provides the frequencyand complementary uses: distribution of V, D, and J region segment utilization using the Illumina MiSeq platform.1. Detection of clonal populations. 2. Identification of sequence information and gene segment utilization. Analysis of the rearranged TRB locus increases the probability 3. Ability to track sequences in subsequent samples with the use of the of identifying T-cell receptor gene rearrangements, as compared Invivoscribe MRD Software. to testing for TRG gene rearrangements only. As a result, combining the analysis of TRB and TRG loci increases the sensitivity The human T-cell receptor beta (TRB) gene locus on chromosome of clonality detection.7 (7q34) includes 65 V (variable) gene segments, followed by two separate clusters of genes each containing a D (diversity) gene, several J (joining) genes, and a C (constant) region spread over Summary and Explanation of the Test 685 kilobases. The two Cgenes, TRBC1 and TRBC2, encode highly This assay utilizes a single multiplex master mix to target conserved homologous products with no functional difference.V, D and J regions of the human TRB gene locus described in lymphoid malignancies, thereby reducing sample DNA requirements andDuring lymphoid cell development, antigen receptor genes simplifying the testing workflow. undergo somatic gene rearrangements. 1Specifically, during T-cell development, genes encoding TRB molecules are assembled The LymphoTrack TRB master mix primers are also designed withfrom multiple polymorphic gene segments that generate V-D-J Illumina adapters and 8 indices (Kit A) or 24 indices (Panel). combinations unique in both length and sequence. 2This allows amplicons generated from different indexed TRB Since leukemias and lymphomas originate from the malignant master mixes to be pooled into a single sequencing library. transformation of individual lymphoid cells, all leukemias and The associated LymphoTrack Software is capable of sorting lymphomas generally share one or more cell-specific or clonal complex NGS data by gene target. This offers a second layer antigen receptor gene rearrangements. Therefore, tests that detect of multiplexing to reduce per sample testing costs by allowingTRB clonal rearrangements can be useful in the study of B- andamplicons from any LymphoTrack Assay (e.g. IGH, IGK, TRB, TRG)T-cell malignancies.to be sequenced on the same flow cell. In addition, the LymphoTrack Software provides easy visualization of data and the LymphoTrackNote: For a more thorough explanation of the locus and the targeted MRD Software allows identified sequences to be tracked anddeep sequencing strategy, please refer to Principle of Immunoglobulin monitored in subsequent samples. and T-Cell Receptor Gene Rearrangement. 2 Positive clonal and negative polyclonal DNA controls are included in kits. Please see the LymphoTrack MRD software section to learn howSpecimen Requirementthe LymphoTrack Assays can be applied to MRD studies, or email 50 ng of high-quality genomic [email protected]. Tonegawa, S. (1983). Nature 302, 575-581.2. Miller JE. (2013) Molecular Genetic Pathology(2nd Edition.,sections 30.2.7.13 and 30.2.7.18).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. 52'