GSM7697199: olfactory epithelium, Ms4a6c KO, rep1; Mus musculus; RNA-Seq
Three 8-12-week old Ms4a cluster knockout, Ms4a6c knockout, and littermate wild-type control mice were euthanized according to our IACUC protocol, and the main olfactory epithelium was dissected and immersed quickly in 0.6 ml of ice-cold Trizol (Invitrogen, #15596018). Olfactory epithelia were ground and homogenized with nuclease-free disposable pestles (Fisher Scientific, #12-141-368) in an Eppendorf tube (Eppendorf, #2231000347). The homogenized sample was incubated for 5 minutes at room temperature. 0.1 µL of chloroform was added to the sample and mixed by inverting for 20 seconds. The sample was then incubated for 3 minutes at RT. The sample was centrifuged at 12,000 xg for 15 minutes at 4 °C. The clear aqueous phase was collected into a nuclease-free Eppendorf tube, equal volume of 70% ethanol (Sigma-Aldrich, #E7023) was added and mixed. The sample was then loaded into RNeasy spin column and RNA was extracted with a RNeasy mini kit (Qiagen, #74104) following the manufacturer's guidelines. NEBNext Ultra II Directional RNA Library Prep Kit for Illumina (E7490S and #E7760S) was used to construct sequencing libraries following the manufacturer's guidelines with one alteration, which was to increase the insert length to around 300 bp.