Oncomine™ BCR IGH LR Assay, RNA - FAQs

The assay provides sufficient reagents for rapid preparation of 24 sample libraries.

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The Oncomine BCR IGH LR Assay, RNA consists of the Oncomine BCR IGH-LR Panel, the Ion AmpliSeq Library Kit Plus (Cat. No. 4488990), and the Ion Torrent NGS Reverse Transcription Kit (Cat. No. A45003). Suffic’ent reagents are provided for the rapid preparation of 24 sample libraries.

Find additional tips, troubleshooting help, and resources within our Next-Generation Sequencing Support Center.

The assay input requirement ranges from 25 ng to 2 µg DNase-treated total RNA. For RNA samples with a concentration of <6.25 ng/µL, we recommend using the DNAclear Purification Kit (Cat. No. AM1756) to concentrate the sample after performing reverse transcription. We recommend using ≥ 50 ng DNase-treated total RNA depending on sample type, ranging from 50-2,000 ng (see "Saturation analysis of peripheral blood leukocytes (PBL) - Oncomine BCR IGH Assay" on page 159 and "Saturation analysis of tissue - Oncomine BCR IGH Assay" on page 161 of the user guide (https://assets.thermofisher.com/TFS-Assets/LSG/manuals/MAN0017438_OncomineTCRBetaAssay_UG.pdf).

When using degraded samples (FFPE) or samples with unknown B or T-cell content, perform sample qualification and quantitation to determine the minimum sample input that is required. For more information, see Appendix B, "Sample qualification and quantitation" on page 130 of the user guide.

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This assay is compatible with isolated B cells, whole blood, peripheral blood leukocytes (PBLs), peripheral blood mononuclear cells (PBMCs), and fresh frozen tissue samples, including bone marrow samples.

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The Oncomine BCR IGH LR Assay, RNA (Cat. No. A45485) is a highly sensitive, RNA-based NGS assay that enables the characterization of immunoglobulin heavy chain sequences, including all complementarity-determining regions (CDR1, 2, and 3) and the CH1 domain of the constant gene. The assay accurately measures B cell repertoire diversity and clonal expansion in a wide array of sample types. It allows for characterization of B cell isotype (and subtype), and the level of somatic hypermutation across the B cell receptor heavy chain.

Find additional tips, troubleshooting help, and resources within our Next-Generation Sequencing Support Center.