GSM5009132: HeLa cells, siTRMT2A rep_2; Homo sapiens; ncRNA-Seq
The total RNA was extracted using the mirVanaTM miRNA Isolation kit (Ambion) or TRIsure™ (Bioline) according to manufacturer´s instructions. The integrity and quantity of each RNA sample were assessed using a Bioanalyzer. Total RNA samples were first pretreated as following to remove some  RNA modifications that interfere with small RNA-seq library construction: 3'-aminoacyl (charged)  deacylation to 3'-OH for 3'-adaptor ligation, 3'-cP (2',3'-cyclic phosphate) removal to 3'-OH for 3'  -adaptor ligation, 5'-OH (hydroxyl group) phosphorylation to 5'-P for 5'-adaptor ligation, m1A and  m3C demethylation for efficient reverse transcription. Then pretreated total RNA was used to prepare  the sequencing library in the following steps: 1) 3'-adapter ligation; 2) 5'-adapter ligation; 3) cDNA  synthesis; 4) PCR amplification; 5) size selection of 134_x0018_160bp PCR amplified fragments (corresponding  to 14_x0018_40nt small RNA size range). The libraries were denatured as single-stranded DNA molecules,  captured on Illumina flow cells, amplified in situ as sequencing clusters and sequenced for 50 cycles on  Illumina NextSeq 500 system per the manufacturer's instructions. Small-RNA Seq