title: T cell receptor beta sequences from human peripheral blood-derived CD4 memory T cells
design: Briefly, cDNA was amplified using the SmartScribe RT kit (Clontech) followed by use of all the cDNA in the first round PCR reactions using primers as specified by Mamedov et al. (40, 41) and Q5 Polymerase (NEB). First round PCR products were purified (Qiagen) to remove excess primers, and a second round PCR performed with nested primers. This product was gel purified (Qiagen) and immediately quantified and used for library preparation using the NEBNext Library kit for Illumina (NEB). Library integrity was checked by TapeStation (Agilent Technologies) analysis followed by quantification using qPCR. The healthy control PB samples and the HealthyControl-PB samples were sequenced separately on Illumina MiSeq paired-end 150bp lanes using separate indexing for ? and ? TCRs by the OMRF Next Generation Sequencing Core.