title: GSM7518763: HUVECs with HG+Mg2+medium-3; Homo sapiens; RNA-Seq
library_construction_protocol: Total RNA was extracted with Trizol reagent (Invitrogen, Carlsbad, CA, USA) and RNA integrity and quantity were measured using the Agilent 2100/4200 system. RNA library for RNA-seq was prepared as fllows: mRNA was purified from total RNA using polyT and then fragmented into 300~350 bp fragments, the first strand cDNA was reverse-transcribed using fragmented RNA and dNTPs (dATP, dTTP, dCTP and dGTP) and second strand cDNA synthesis was subsequently performed. Remaining overhangs of double-strand cDNA were converted into blunt ends via exonuclease/polymerase activities. After adenylation of 3' ends of DNA fragments, sequencing adaptors were ligated to the cDNA and the library fragments were purified. The template was enriched by PCR, and the PCR product was purified to obtain the final library.