GSM5005597: P201_1; Mus musculus; RNA-Seq
Total RNA extraction was performed using RNeasy Mini Kit. RNA was eluted using Nuclease-Free water and total yield and purity of RNA were assessed using NanoDrop™ 2000 (Thermo Fisher). RNA from cells quantified using the Quant-iTTM RiboGreen® RNA Assay Kit and normalized to 5 ng/μL. An automated variant of the Illumina TruSeqTM Stranded mRNA Sample Preparation Kit was used for library preparation from a 200 ng aliquot of RNA. This method preserves strand orientation of the RNA transcript and uses oligo dT beads to select mRNA from the total RNA sample. Following cDNA synthesis and enrichment, cDNA libraries were quantified with qPCR using KAPA Library Quantification Kit for Illumina Sequencing Platforms and then pooled equimolarly.