GSM8502608: WT BR2; Mus musculus; OTHER
Live CD4+CD25- TCRb+, aGalCerCd1d tetramer- cells were sorted with purity of 98% or higher. RNA was isolated using the RNeasy Plus Mini Kit (Qiagen). In order to analyze TCR repertoire, we used the SMARTer mouse TCR a/b profiling kit (Takara, cat no 634403) following the manufacturer's instructions. We used 20 ng of total RNA  in order to synthesize cDNA for each library. The kit employs a 5'-RACE-based approach to capture complete V(D)J variable regions of TCR transcripts. The libraries were sequenced using a Miseq sequencer (Illumina) at the UNC High Throughput Sequencing Facility.  The alignment and assembly of the RNAseq reads and the exporting of the clonotypes were conducted using MiXCR.