GSM5068353: Control_replicate_1; Mus musculus; RNA-Seq
Freshly dissected olfactory epithelia were minced to ~1mm3 cubes with a razor blade on a glass slide on ice, and placed in a scintillation vial for dissociation. To isolate individual cells we used the Papain Dissociation System (catalog # LK003150, Worthington Biochemical Corporation, Lakewood, NJ), with the following modifications: 1) we incubated olfactory epithelia in papain solution for only 15 minutes versus the recommended time without an appreciable loss of cells, 2) after trituration, the cell suspension was filtered through a 40 μm mesh strainer (catalog # 352340, BD Bioscience, San Jose, CA). Dissociated cells were resuspended in 1ml ice-cold 1x Hank's balanced salt solution (HBSS) (catalog # 14025 , Life Technologies, Grand Island NY) and then sorted. Each OE was prepared and sorted within 1.5 hours. Samples were visually inspected under a microscope.  Illumina TruSeq RNA sample preparation protocol