title: GSM7964845: siNC_1; Homo sapiens; RNA-Seq
library_construction_protocol: Total RNA was treated with RQ1 DNase (Promega) to remove DNA. The quality and quantity of the purified RNA were determined by measuring the absorbance at 260 nm/280 nm (A260/A280) using Nanodrop One (Thermo). RNA integrity was further verified by 1.5% agarose gel electrophoresis. For each sample, 1 μg of total RNA was used for RNA-seq library preparation using KAPA Stranded mRNA-Seq Kit for Illumina® Platforms (#KK8544). Polyadenylated mRNAs were purified by VAHTS mRNA capture Beads (N401-01), and fragmented mRNAs were converted into double-strand cDNA. Following end repair and A tailing, the DNAs were ligated to Diluted Roche Adaptor (KK8726). After the purification of ligation product and size fractioning to 300–500 bps, the ligated products were amplified, purified, quantified, and stored at −80 °C before sequencing. For strand-specific sequencing, the strand marked with dUTP (second cDNA strand) was not amplified. For high-throughput sequencing, the libraries were applied to Illumina Novaseq 6000 system for 150 nt paired-end sequencing.