TCR NORMAL DONOR LONGITUDINAL SAMPLE: Sample HEALTH CONTROL_S3_1
A total of 2 million peripheral mononuclear cells (PBMCs) were used from each HAM/TSP patients and healthy controls to generate a TCR-ß library. Total RNA was extracted using the RNeasy plus kit (Qiagen, USA) and cDNA synthesis was performed using the anchored switch 5’rapid amplification of cDNA ends (5’RACE) PCR-based primer combined with a Unique Molecular Identifier (UMI) that consists of 10 random nucleotides sequence (AAGCAGTGGTATCAACGCAGAGTACNNNNNNNNNNUCTTrGrGrGrG). The human TCR V-D-J ß-chain region was amplified using a pair of primers specific for the TCR constant region (TGCTTCTGATGGCTCAAACACAGCGACCT) and 5’RACE region.
alignment_software
MiGEC