GSM9772082: HDLEC, non-silencing siRNA control, biological replicate 3; Homo sapiens; RNA-Seq
RNA extraction was performed using RNeasy Mini Kit (Cat No. 74104, QIAGEN). RNA concentration was quantified using Nanodrop and all samples were deemed pure if  280/260 ratio > 1.75. RNA samples were diluted to 5µg using RNAase free water.  First strand cDNA was synthesized using random hexamer primer 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. 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 (Beckman Coulter, USA). Transcriptomes were read by Illumina HiSeq platforms by Novogene Inc (Hong Kong, China) in a paired-end manner to a depth of 20M reads.