The Oncomine IGH FR3(d)-J Assay is a targeted next-generation sequencing (NGS) assay for detection of B-cell diversity to measure clonal expansion/contraction with accuracy and for detection of rare clones with high sensitivity. Primers have been specifically designed to target the distal region of FR3 to J of the immunoglobulin heavy chain of the B-cell receptor. Due to lower levels of somatic hypermutation observed in the distal region of FR3, there is an increase in primer-binding efficiency. When combined with Ion Reporter Software, data can be easily analyzed and reports run within a short period of time.

Key features of the Oncomine IGH FR3(d)-J Assay:
• Sequencing of FR3(d)-J region of IGH chain
• Unique primer design to distal FR3 region may reduce primer mismatches due to somatic hypermutation
• Detect rare clones at LOD of 10^-6
• Optimized Oncomine informatics for easy, all-inclusive analysis and publication-ready data plots

The kit provides reagents for library construction and dNTPs. Ion AmpliSeq libraries can be generated from DNA extracted from FFPE tissue, fresh/frozen tissue, bone marrow, whole blood, peripheral blood leukocytes, PBMCs, and sorted B cells with as little as 50 ng of nucleic acid. The assay is optimized for sequencing on the Ion GeneStudio S5 System with the Ion 530, Ion 540, and Ion 550 chips to enable up to 24 samples per run.

When the primary test isn’t enough
Often when the desired results are not achieved with a primary test, an assay designed with primers directed to different regions of the V(D)J region can be used as a secondary or follow-up test to improve sensitivity of detection. Where needed, secondary testing of samples can be achieved using the Oncomine IGH FR3(d)-J Assay.