GSM6895379: iTreg_48h_LIRIL2R_LNA1_rep3; Homo sapiens; RNA-Seq
RNA was isolated (RNeasy Mini Kit, cat# 74106, QIAGEN) and treated in-column with DNase (RNase-Free DNase Set; QIAGEN) for 15 min. The removal of genomic DNA was ensured by an additional treatment of the samples with DNase I (Invitrogen). RNA quantification was performed using Nanodrop 2000 and quality control was performed using BioRad Experion or Agilent Bioanalyzer. Libraries for RNA-Seq were prepared using Illumina TruSeq® Stranded mRNA Sample Preparation Guide (part #15031047). The quality of the libraries was confirmed with Advanced Analytical Fragment Analyzer, and the concentrations of the libraries were quantified with Qubit® Fluorometric Quantitation (Life Technologies). 24h samples were sequenced togother in two lanes. Each sample was sequenced on both lanes. 48h and 72h samples were sequenced togther in one lane. Samples for 24h were prepared at different times as compared to 48h and 72h, which were prepared from the same culture. Sequencing was performed at the Finnish Functional Genomics Centre using the HiSeq3000 Next-Generation Sequencing platform.