GSM8878111: ECEV_72h_2; Homo sapiens; RNA-Seq
RNA was harvested using Trizol and purified with an RNA Clean & Concentrator-25 kit (Zymo, Tustin, CA, USA). RNA samples were submitted to Novogene (Novogene Corporation Ince., Sacramento, CA, USA) for quality control, library preparation, sequencing, and quantitation as per the in-house standard for gene expression analysis. The library preparation, genome mapping, and raw gene counts for mRNA-sequencing analysis were performed by Novogene (Novogene America). The following methods were those provided by Novogene. Libraries generated for mRNA-sequencing were checked with Qubit and real-time (RT) PCR for quantification and bioanalyzer for size distribution detection. Quantified libraries were pooled and sequenced on Illumina sequencing platforms (Illumina, Omaha, NE, USA) and clean data was obtained by using in-house Perl scripts from Novogene (Novogene America, Sacramento, CA, USA). Hisat2 (v2.0.5) was used to index and map reads to the human reference genome (GRCh38/hg38), and the mapped reads were assembled by StringTie (v1.3.3b) [22]. Read numbers mapped to each gene were counted in R using the FeatureCounts package (v1.5.0-p3) and an FPKM of each gene was calculated [23].