GSM9765057: HUVEC, non-silencing siRNA control, biological replicate 1; Homo sapiens; RNA-Seq
RNA extraction was performed using RNeasy Mini Kit (Cat No. 74104, QIAGEN). Libraries were prepared by Novogene. RNA samples were diluted to 5µg using RNAase free water. The first strand cDNA was synthesized using random hexamer primers and M-MuLV Reverse Transcriptase (RNase H-). Second strand cDNA synthesis was subsequently performed using DNA Polymerase I and RNase H. Double-stranded cDNA was purified using AMPure XP beads. Remaining overhangs of the purified double-stranded cDNA were converted into blunt ends via exonuclease/polymerase activities. After adenylation of 3' ends of DNA fragments, NEBNext Adaptor with hairpin loop structure was ligated for hybridisation. cDNA fragments of 150-200bp in length were generated by PCR and purified using the AMPure XP system