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Protocol for high-throughput genome-wide translocation sequencing-based profiling of T-cell receptor α and β gene
Rui Luo1, Yawei Song2, Dingpeng Yang3
1First Affiliated Hospital of Chongqing Medical University, Chongqing Medical University, Chongqing 400016, China; State Key Laboratory of Epigenetic Regulation and Intervention, Shanghai Institute of Biochemistry and Cell Biology, Center for Excellence in Molecular Cell Science, Chinese Academy of Sciences, University of Chinese Academy of Sciences, Shanghai 200031, China.
Abstract:
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.
