RNA extraction from TRIzol cell lysate was performed according to the manufacturer’s manual. mRNA purified from extracted total RNA with Dynabeads mRNA DIRECT Kit (Life Technologies) was used for reverse transcription, during which the 5’end of TCRα mRNA was tagged with a synthesized oligo bearing random barcodes AAGCAGTGGTATCAACGCAGAGUNNNNUNNNNUNNNNUCTTrGrGrGrGrG (N for random nucleotides, r for RNA) via template switch. Sample indices and sequencing adaptors were introduced during the PCR amplification of the cDNA. Libraries were quantified with KAPA Library Quantification Kits. Indexed samples were mixed and sequenced with Illumina MiSeq Reagent Kit V3 for paired-end 200 + 200 cycles.
GEO Accession
GSM1828722