GSM2575443: 2.6 Rod+; Mus musculus; RNA-Seq
For RNA-seq Total RNA was extracted using the RNeasy kit (Qiagen) ~2 ng of total RNA samples (range 0.4-5 ng) were processed with the SMART-Seq v4 Ultra Low Input RNA Kit (Clontech) following the manufacturer instructions. Resulting cDNA was sheared on a S220 Focused-ultrasonicator (Covaris) and subsequently processed with the "NEBNext Ultra II DNA Library Prep Kit for Illumina" (NEB #E7645). Briefly, oligo(dT)-primed reverse transcription was performed in presence of a template switching oligonucleotide, double stranded cDNA was produced by 14 cycles of PCR and submitted to acoustic shearing. Fragments were processed through subsequent enzymatic treatments of end-repair, dA-tailing, and ligation to Illumina adapters. Adapter-ligated libraries were completed by limited-cycle PCR (8 cycles).