b"11.What are the differences between the TCRG Gene Clonality Assay and the T-Cell Receptor Gamma Gene Rearrangement Assay 2.0?15.What are the recommended settings for my ABI instrument?The TCRG Gene Clonality Assay was designed by the EuroClonality/ BIOMED-2 Group and consists of two masterInstruments should be calibrated with the DS-30 matrix mixes. For polyclonal populations, four Gaussian distributions arestandards (Dye set D) for the ABI 310, 3100, or 3130 instrument generated. The T-Cell Receptor Gamma Gene Rearrangementseries. For the ABI 3500 sequencer series, we advise that you Assay 2.0 was designed by Invivoscribe and it's performancecalibrate the instrument with DS-33 matrix standards. We also was subsequently reviewed and validated by the EuroClonality/ recommend using either POP-4 or POP-7 depending on which BIOMED-2 Group. It targets all functional V H- J HrearrangementsABI instrument you are using. If your equipment supports POP-7, in a single master mix and produces smaller ampliconswe recommend using this polymer as it can be utilized for both grouped under a single Gaussian distribution. This allows forfragment analysis and sequencing; whereas, POP-4 can only be easier interpretation and makes the assay more suitable forutilized for fragment analysis.DNA extracted from FFPE tissue, which may consist of partially 16.How should peaks outside the valid size range be interpreted degraded DNA that would not amplify well with the larger valid size range of the TCRG Gene Clonality Assay. when using assay kits?12.What are the differences between the IGH-BCL2 TranslocationYou should not interpret peaks outside of the valid size range; although, in theory, it is possible to have a true rearrangement Assay and the IGH-BCL2 t(14;18) Translocation Assay? fall outside this region. If you are concerned about a suspect The IGH-BCL2 Translocation Assay was designed by thepeak, you may sequence your product for confirmation. Please EuroClonality/BIOMED-2 Group and is available as either anote that samples should always be interpreted within the CE-IVD or research use only assay whereas; the IGH-BCL2context of all available clinical information.t(14;18) Translocation Assay was designed by Invivoscribe 17.Is cell-free DNA (cfDNA) a suitable sample type for Invivoscribe and is only available for research use only. Both of these assays target MBR and Mcr translocations, but the IGH-BCL2LymphoTrack or LymphoTrack Dx Assays? Translocation Assay also targets the translocations at the 3The average size of cfDNA (~170 bps) makes it a suitable sample Mbr. The IGH-BCL2 t(14;18) Translocation Assay was type to run with IGH FR3 master mixes. The use of cfDNA with designed as a nested PCR allowing greater sensitivities TRG master mixes might be possible, but expected amplicon (1 clonal cell per 10,000 normal cells) to be achieved. The limitsizes generated with this assay are near the upper limits of the of detection of the IGH-BCL2 Translocation Assay is 1 clonalfragment lengths typically found with this sample type.cell per 100 normal cells. Lastly, the IGH-BCL2 Translocation Assay includes 33 reactions, whereas the IGH-BCL2 t(14;18)18. s DNA extracted from FFPE tissue suitable to use with ITranslocation Assay includes 30 reactions. Invivoscribe LymphoTrack or LymphoTrack Dx Assays? 13.Do you offer quantitative chromosome translocation To ensure DNA from challenging specimens is of sufficient quality and quantity to generate a valid result, samples may be (e.g., BCR-ABL1) controls? tested with the Specimen Control Size Ladder master mix. Our controls are validated for qualitative use, although our customers do successfully use them with quantitative assays.19.On which instruments can I use the LymphoTrack andUnfortunately, we cannot guarantee their performance with anyLymphoTrack Dx Assays?assay that was not designed by Invivoscribe. We have different versions of our assays for the S5/PGM and 14.Which capillary electrophoresis instruments are currentlyMiSeq instruments (LymphoTrack TRB is currently available only on MiSeq). No other DNA sequencers (e.g. 454) are currently validated for use with our assay kits? supported. Assays for the Ion S5/PGM and MiSeq platforms Currently the capillary electrophoresis instruments Invivoscribediffer slightly in terms of the total number of indices, etc., but has validated include: ABI 3100 and 3130 series for all capillaryboth have similar benefits such as a one-step PCR reaction and electrophoresis detection assays. The ABI 310 and 3500included bioinformatics software.instrument series have also been validated for the majority of our capillary electrophoresis detection assays. We are not20.and How much DNA is needed for the LymphoTrackable to support using instruments not listed as validated in theLymphoTrack Dx Assays?instructions for use of our CE-IVD assays.50 ng of high-quality genomic DNA is required for the Ion S5/PGM and MiSeq LymphoTrack and LymphoTrack Dx Assays for clonality and somatic hypermutation applications.Invivoscribe 2020|159"