GSM8615924: F11_HC_subject2_PDC-E2_Tconv; Homo sapiens; RNA-Seq
To optimize recovery, PDC-E2-reactive T cells were sorted 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 Bio) as previously described 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 NextSeq 2000 sequencer (Illumina) with 58-base reads, using HiSeq v4 Cluster and SBS kits (Illumina) with a target depth of 1 million reads per sample.