title: GSM4409011: 10_Standard culture, Basolateral infection (MOI 0.7), Replicate 1; Homo sapiens; RNA-Seq
library_construction_protocol: Cell lysates were stored in liquid nitrogen before RNA isolation. Isolation of total RNA from ECs was conducted using the ZR-Duet DNA/RNA MiniPrep Plus kit (Zymo Research, Irvine, CA, USA) according to manufacturer's instructions. RNA samples were digested using DNase I and RNA integrity was accessed using automated capillary electrophoresis (RNA pico sensitivity assay, LabChip GX II Touch HT, Perkin Elmer, Villebon-sur-Yvette, France). Genome-wide gene expression profiling was performed using the MACE-seq method. In general, the procedure follows a modified protocol described in Nold-Petry et al. (2015). In brief, samples with 100 ng of DNase-treated RNA were used for library preparation. Synthesis of cDNA was performed by reverse transcription using oligo (dT) primers following fragmentation of cDNA to an average size of 200 bp using sonification (Bioruptor, Diagenode, Seraing, Belgium). DNA was quantified using a Qubit HS dsDNA assay (Thermo Fisher Scientific). cDNA fragments were ligated to DNA adapters containing TrueQuant unique molecular identifiers included in the kit. Library amplification was done using polymerase chain reaction, purified by solid phase reversible immobilization beads (Agencourt AMPure XP, Beckman Coulter, Brea, CA, USA).
attributes.tag: GEO Accession
attributes.value: GSM4409011