GSM8323147: Nuc_Control_3; Mus musculus; RNA-Seq
Cellular fractionation was performed to isolate cell nucleus and immunoprecipitation was performed as described previously (Vangoor et al., JNeu. 2019). Three samples per group were used for RNA sequencing. The NEBNext Ultra Directional RNA Library Prep Kit for Illumina was used to process the samples. Sample preparation was performed according to NEBNext Ultra Directional Library Prep Kit for Illumina (NEB #E7420S/L) protocol. Briefly, by using the oligo-dT magnetic beads mRNA was isolated from total RNA, followed by fragmentation of the mRNA and cDNA synthesis. The resulting cDNA was ligated with the sequencing adapters and PCR amplification. The yield and quality of the samples was estimated using a Fragment analyzer, where the size of the resulting products was consistent with the size distribution showing a broad peak between 300 – 500 bp. Subsequently, clustering and DNA sequencing was performed using the Illumina cBot and HiSeq 4000, according to the manufacturer's protocol. HiSeq control software HCS v3.3.52 was used. The sequencing experiments were performed at GenomeScan B.V., Leiden, The Netherlands.