GSM6394812: 3_Trans_NoStim [lib27726]; Homo sapiens; RNA-Seq
The indicated populations were isolated by flow cytometry, and after stimulation cells were resuspended in SMARTer v4 lysis reagents (Clontech). Cells were lysed and cDNA was synthesized. For the cell lines, stimulated or unstimulated cells were used for RNA extraction with QIAGEN RNeasy Mini kit and cDNA synthesized from this. After amplification, sequencing libraries were prepared using the Nextera XT DNA Library Preparation Kit (Illumina). Barcoded libraries were pooled and quantified using a Qubit® Fluorometer (Life Technologies). Single-read sequencing of the pooled libraries was carried out on a HiSeq2500 sequencer (Illumina) for 74 cycles, using TruSeq v3, and SBS kits (Illumina). Target read depths were ~5-10 million raw reads per sample.