Protocol for high-throughput genome-wide translocation sequencing-based profiling of T-cell receptor α and β gene rearrangements in mouse T cells.

Current T cell receptor (TCR) sequencing approaches have inherent technical limitations, and low-primer-bias techniques for quantitative genomic-level repertoire analysis remain limited. Here, we present high-throughput genome-wide translocation sequencing-based TCR sequencing (HTGTS-TCR-seq) as a genomic DNA-based protocol for low-bias analysis of TCR diversity. We describe steps for DNA fragmentation, LAM-PCR enrichment, streptavidin bead capture, adapter ligation, and PCR for library construction. This protocol enables quantitative profiling of TCR rearrangement products at the genomic DNA level. For complete details on the use and execution of this protocol, please refer to Luo et al.1.