title: GSgt10012
design: none provided
library_construction_protocol: RNA was extracted using Isogen protocol and cDNA synthesized using the  SuperScript II kit (Invitrogen) with oligo-dT primers (Invitrogen)  according to manufacturer?s protocol. cDNAs were then amplified using  ExTaq polymerase kit (TAKARA). TRAV14 (CCAATCTCTGACAGTCTGGGAA) and TRAC  (AACTGGTACACAGCAGGTTCTG) primers were used for TRAV14 repertoire; TRBV2  (CGAACAGTATCTAGGCCACAATGC) and TRBC (GCACACGAGGGTAGCCTTTTGTTT) primers  for TRBV2 repertoire. PCR program was as follow: 4 min at 94 ?C; x  cycles of 30 s at 94 ?C, 30 s at 62 ?C, 30 s at 72 ?C; 5 min at 72 ?C 5;  hold at 4 ?C. The number of cycles was adapted to the initial number of  cells (24 or 25 cycles for conventional T cells, 27 to 29 for regulatory  T cells) as we observed the formation of heteroduplexes in saturated  reactions. PCR products were sequenced on a Roche Genome Sequencer FLX  Titanium system after adapter ligation following standard procedures.