Human TRB repertoires of children with CHD and aged-matched controls
mRNA extraction was performed from thawed FACS-sorted cells according to instructions of the RNeasy mini kit (Qiagen). For TRB repertoire analysis, the isolated mRNA was reverse-transcribed using SMARTer RACE 5-3 PCR Kit (Clontech) with a customized protocol. Next, the CDR3 region of TRB clones was amplified via RACE PCR. For all conditions and to control PCR contamination, negative controls (H2O) were run together with each PCR, and also subjected to sequencing. All PCR amplicons were subsequently prepared for paired-end Illumina sequencing (v2 500 cycles), using the Illumina MiSeq platform according to the Illumina guidelines.