GSM8006517: Cord blood, rep4; Homo sapiens; RNA-Seq
Total RNA was extracted with RNeasy plus micro kit (Qiagen) following manufacturer's instructions and eluted in 14 µL of RNase free water 1-10 ng purified total RNA was converted into cDNA and amplified (9-12 cycles) using the SMART-Seq HT kit (Takara Bio) or the SMART-Seq v4 Ultra Low Input RNA kit (Takara Bio), for Tfr analyzed ex vivo or after the in vitro differentiation, respectively. 10 ng of cDNA were used to prepare DNA libraries using Nextera XT DNA Library Preparation Kit (96 samples, Illumina) and Nextera XT Index Kit (96 Indexes, Set A, Illumina). Next, Barcoded samples were pooled, diluted at 1.8 pM and loaded onto a NextSeq 500/550 High Output Kit v2.5 flow cell (75 Cycles, 120-300 million of reads, Illumina). Single-end sequencing was performed on a NextSeq 550 (Illumina).