GSM5668613: DeltaLN_siCtl_2; Mus musculus; RNA-Seq
Total RNA was isolated from the samples above using the miRNeasy Mini Kit (Qiagen, 217004) with on-column DNAse digestion according to the manufacturer's instructions. 5g of RNA was then subjected to rRNA depletion using the Ribozero magnetic gold kit (Epicentre, MRZG12324) followed by concentrating it using the  RNeasy MinElute Cleanup Kit (Qiagen, 74204). Total RNA was isolated from the samples above using the miRNeasy Mini Kit (Qiagen, 217004) with on-column DNAse digestion according to the manufacturer's instructions. RNA quality control was performed with an RNA ScreenTape on the Agilent 4200 TapeStation and the concentration measured by using the Quanti-iT RiboGreen RNA assay Kit (Life Technologies). 200 ng of RNA were subjected to rRNA depletion and utilized for library preparation with the Truseq Stranded Total RNA Library Prep kit with Ribo-Zero Gold (Illumina). Library QC was performed with a Fragment Analyzer (AATI) using the Standard Sensitivity NGS Fragment Analysis Kit (DNF-473). RNA-seq libraries were sequenced SR81 with NextSeq 500 High Output v2 kit (Illumina) on an Illumina NextSeq 500 using protocols defined by the manufacturer; primary data analysis was done using Illumina RTA Version: 2.4.11.