GSM5702811: TCR-seq of homo sapiens:HD12_S4_alpha; Homo sapiens; RNA-Seq
Complementarity determining region (CDR) 3 sequences of TCR alpha and beta chains were amplified by using a modified RACE approach Total RNA extracted from WT1-specific T cell cultures was reverse transcribed and amplified using primers specific for α/β chain constant regions and for the template switching sequence added during cDNA synthesis. A double barcoded strategy was used: each individual sample was unequivocally tagged with a barcoded fusion primer specific for the constant TCR genes in addition to the barcoded template switching primer used during cDNA synthesis. PCR amplicons were purified using the AmPure beads (Beckman Coulter) and sequenced with the Illumina MiSeq_600v3. Illumina MiSeq_600v3: paired end