title: GSM4408210: A549_APEX1_2nd; Homo sapiens; RNA-Seq
library_construction_protocol: Total RNA was isolated from A549 cells with Trizol. The RNA was further purified with two phenol-chloroform treatments and then treated with RQ1 DNase. Purified RNA was transcribed for cDNA by oligo dT primer. And then the insert fragment was synthesized by PCR amplification. A linearized vector digested by EcoRI and XhoI (NEB) and PCR insert were added to a PCR microtube for ligation with ClonExpress® II One Step Cloning Kit (Vazyme). The quality and quantity of the purified RNA were redetermined by measuring the absorbance at 260 nm/280nm (A260/A280) using Smartspec Plus (BioRad, USA). The integrity of RNA was further verified by 1.5% agarose gel electrophoresis. For each sample, 1μg of the total RNA was used for RNA-seq library preparation by VAHTS Stranded mRNA-seq Library Prep Kit (Vazyme). Polyadenylated mRNAs were purified and fragmented, and then converted into double strand cDNA. After the step of end repair and A tailing, the DNAs were ligated to VAHTS RNA Adapters (Vazyme). Purified ligation products corresponding to 200-500 bps were digested with heat-labile UDG, and the single strand cDNA was amplified, purified, quantified and stored at -80℃ before sequencing. For high-throughput sequencing, the libraries were prepared following the manufacturer's instructions and applied to Illumina HiSeq X Ten system for 150 nt paired-end sequencing.
attributes.tag: GEO Accession
attributes.value: GSM4408210