title: GSM9216749: siCTR_repl3; Homo sapiens; RNA-Seq
library_construction_protocol: Total RNA was extracted using the NucleoSpin™ RNA Columns (Macherey-Nagel™, DE; cat# 740955), according to manufaturer's instructions. RNA integrity was verified by an Agilent 2100 Bioanalyzer. 1 microgr of RNA was used for the library construction. Messenger RNA was enriched using oligo(dT) magnetic beads. A strand-specific RNA sequencing library was constructed following the BGI Standard mRNA-seq Library Prep protocol. Briefly, the enriched mRNA was fragmented and reverse-transcribed into first-strand cDNA using random hexamer primers. Second-strand cDNA was synthesized using dUTP instead of dTTP to achieve strand specificity. Double-stranded cDNA was purified, end-repaired, adenylated at the 3' ends, and ligated to BGI's proprietary duplex adapters. The adapter-ligated fragments were purified and size-selected (typically for insert sizes of ~200-300 bp). The libraries were then amplified via PCR, and the final products were purified. Library quality and concentration were assessed using a Bioanalyzer and qPCR. The amplified fragmented cDNA of 100 bp in size were sequenced in paired-end mode using the Genome Analyzer (Illumina) with a read length of 100 bp.