GSM7954979: 1_DR0401_Day 60_lib43350; Homo sapiens; RNA-Seq
CD4+CD154+Tetramer+ cells were sorted by BD FACSAria Fusion at single-cell purity directly into individual wells of a 96-well PCR plate with 5μl reaction buffer from the SMART-Seq v4 Ultra Low Input RNA Kit for Sequencing (Takara). Index sorting was performed, thus the phenotype of surface staining could be traced and confirmed for each individual cell. Reverse transcription was performed, followed by PCR amplification to generate full length amplified cDNA. Sequencing libraries were constructed using the NexteraXT DNA sample preparation kit (Illumina) to generate Illumina-compatible barcoded libraries. Libraries were pooled and quantified using a Qubit® Fluorometer (Life Technologies). Dual-index, single-read sequencing of pooled libraries was carried out on a HiSeq2500 sequencer (Illumina) with 58-base reads, using HiSeq v4 Cluster and SBS kits (Illumina) with a target depth of 1 million reads per sample.