RNA Sequencing

Complete transcriptome analysis of DCs isolated from three young and three old individuals were carried out (description of cohort in Supplementary Table 1). RNA isolation was performed using Qiagen RNeasy plus mini kit (Cat#74134) according to the manufacturer’s instructions. The quantity of RNA was in picograms therefore SMARTer® Stranded Total RNA-Seq Kit t with Picogram input was used to construct the libraries (Clontech, Cat #634411). Libraries were quantified and normalized using the Library Quantification Kit from Kapa Biosystems and sequenced as paired-end 100 bp reads on the Illumina HiSeq 2500 platform. Library preparation and all transcriptome [High throughput sequencing (NGS) experiments were performed at UCI Genomic High Throughput Facility, GHTF (http://dmaf.biochem.uci.edu] using Illumina Genome Analyzer which generated about 10–20 million reads (30nt) from DCs derived from young and aged individuals.