title: bulkTCR-seq of Homo sapiens - peripheral blood
design: Four replicates were sequenced for each timepoint. Briefly, CD3+ T     cells were immunomagnetically isolated from thawed PBMCs samples (using anti-CD3     microbeads, Miltenyi), RNA was isolated (Qiagen), and bulk RNase H-dependent     PCR-enable T-cell receptor-sequencing (rhTCR-seq) was performed. This method     utilizes RNase H-dependent PCR to enhanced specificity of TCR amplification, and     incorporates unique molecular identifiers to each cDNA molecule to improve the     accuracy of clonotype frequency estimations. Sequencing was performed on an Illumina     NovaSeq, and then MiXCR was used to assemble TCR-beta     clonotypes.