title: GSM3181634: HUVEC polyA RNA-Seq; Homo sapiens; RNA-Seq
library_construction_protocol: Total RNA was isolated from HUVEC cells using RNeasy mini kit (Qiagen) according to the manufacturer's instructions. Total RNA isolated was quantified and assessed for quality using Agilent BioAnalyzer. High quality DNase treated-total RNA from HUVECs was used for library preparation using TruSeq stranded mRNA library preparation kit from Illumina and sequenced on HiSeq2500 according to the manufacturer's guidelines. Briefly, mRNA enrichment from total RNA was carried out by polyA tail capture using oligo-DT beads prior to heat-based fragmentation and elution of mRNA. The eluted fragments were converted to cDNA and their 3' ends were adenylated. Adapters consisting of index sequences were then ligated to the adenylated cDNA, prior to limited cycle PCR amplification of the adapter ligated cDNA fragments. Amplified libraries were size-selected, clustered and sequenced after analyzing the quality of the library using BioAnalyzer. Quality checked library was sequenced using Illumina HiSeq 2500 in the high output mode to obtain 100bp paired-end reads.