title: GSM3985894: SerE7; Mus musculus; OTHER
library_construction_protocol: Following odor stimulation, the mouse was sacrificed, and the dissected olfactory epithelium was processed for RNA library in reference to the method described by Jiang et al. with some modifications (Jiang et al., 2015).  Pieces of freshly dissected olfactory epithelium were homogenized and centrifuged. 6 μl of a monoclonal Phospho-S6 Ribosomal Protein (Ser240/244) (D68F8) XP Rabbit mAb (Cell Signaling Technologies) was added to the supernatant, and incubated with rotation for 1.5 hours at 4 °C. Protein A Dynabeads (Invitrogen) were added to the antibody mixture and incubated for another hour with rotation at 4 °C. After incubation and washing, the bead-captured RNA was eluted and purified with RNeasy Micro kit (Qiagen). 10 ng of starting RNA was used to make cDNA libraries with the SMART-Seq v4 kit (Clontech) with 11 cycles of PCR. cDNA was purified with AMPure XP beads (Beckman Coulter) and 0.5 ng was used to make cDNA libraries using the Nextera XT DNA Library Preparation Kit (Illumina). 50 base sequencing was performed on the HiSeq 2500 System (Illumina), obtaining 15-38 million reads per sample.
identifiers.namespace: GEO
identifiers.id: GSM3985894