GSM8252898: Ad_Shox2_1; Homo sapiens; RNA-Seq
Total RNA of human vascular smooth muscle cells were extracted using TRIzol regents and RNA quality was determined using Agilent 2100 Bioanalyser. mRNA molecules were purified from total RNA using oligo (dT)-attached magnetic beads. Then, mRNA molecules were fragmented into small pieces using fragmentation reagent after reaction a certain period in proper temperature. First-strand cDNA was generated using random hexamer-primed reverse transcription, followed by a second-strand cDNA synthesis. The synthesis cDNA was subjected to end-repair and then was 3'-adenylated. Adapters were ligated to the ends of these 3'-adenylated cDNA fragments. PCR was used to amplify the cDNA fragments with adapters from previous step. PCR products were purified with Ampure XP Beads (AGENCOURT), and dissolved in EB solution. Library was validated on the Agilent Technologies 2100 Bioanalysers.