GSM8049070: CVN2-1__rep1; Homo sapiens; RNA-Seq
Total RNA from Total RNA from mock or virus-infected calu-3 cells was extracted using the Zymo Direct-zol RNA micro prep Kit. Concentration and integrity (RIN) of isolated RNA were determined using the Quant‐iT™ RiboGreen™ RNA Assay Kit (Thermo Fisher) and an RNA Standard Sensitivity Kit (DNF‐471, Agilent Technologies, Santa Clara, CA, USA) on a Fragment Analyzer Automated CE system (Agilent Technologies), respectively. Subsequently, cDNA libraries were constructed from total RNA using the Universal Plus mRNA‐Seq kit (Tecan Genomics, San Carlos, CA, USA) in a Biomek i7 Automated Workstation (Beckman Coulter). Briefly, mRNA was isolated from purified 300ng total RNA using oligo‐dT beads and used to synthesize cDNA following the manufacturer's instructions. The transcripts for ribosomal RNA (rRNA) and globin were further depleted using the AnyDeplete kit (Tecan Genomics) prior to the amplification of libraries. Library concentration was assessed fluorometrically using the Qubit dsDNA HS Kit (Thermo Fisher), and quality was assessed with the HS NGS Fragment Kit (1–6,000bp; DNF‐474, Agilent Technologies).