title: GSM5013464: Proliferative HASMCs rep1; Homo sapiens; ncRNA-Seq
library_construction_protocol: RNAsimple Total RNA Kit was used to extract total RNA from proliferative and quiescent HASMCs. In order to eliminate the interference of RNA modification on the construction of small RNA-seq library, the total RNA samples were first pretreated as follows: 3ʹ-aminoacyl (charged) was deacylated to 3ʹ-OH for 3ʹ adaptor ligation, 3ʹ-cP (2ʹ, 3ʹ-cyclic phosphate) was removed to 3ʹ-OH for 3ʹ adaptor ligation, 5ʹ-OH (hydroxyl group) was phosphorylated to 5ʹ-P for 5ʹ-adaptor ligation, m1A and m3C were demethylated for effective reverse transcription. Subsequently, the commercial kit for tRF & tiRNA sequencing library preparation was used to construct the tsRNA-seq libraries. The kit includes 3ʹ-adapter and 5ʹ-adapter ligation adaptor ligation, cDNA synthesis and library PCR amplification. PCR amplified fragments with a size of 135~160 bp (corresponding to the size range of 15~40 nt small RNA) were selected as tsRNA-seq libraries. Finally, the prepared tsRNA-seq libraries were quantified using Agilent BioAnalyzer 2100, and then sequenced using Illumina NextSeq 500.