title: RNA-seq of Human Brain Microvascular Endothelial cells
design: RNA was extracted using TRIzol from HBMEC, RNA concentration was confirmed by NanoDrop. cDNA library construction was performed following BGISEQ-500 platform; mRNA enriched by Oligo (dT) magnetic beads, then target RNA was fragmented, and N6 random primers were used for dscDNA synthesis by reverse transcription. dscDNA were subjected to end repair and 3 end adenylated, Adaptors were ligated at the 3 end and the PCR was  amplified using specific primer. Furthermore, the PCR product was denatured into single stranded DNA and cyclized with splint oligo and DNA ligase, DNB was prepared and sequenced for SE50.